METHODS AND COMPOSITIONS FOR SCREENING FOR OR MODULATING A TUMOR ASSOCIATED ANTIGEN

Abstract: Methods of screening for or treating cancer are disclosed. The screening methods are based on the detection of an antigen, or a nucleic acid molecule encoding the antigen, found by the present invention to be associated with the presence of cancer. Preferred embodiments of the methods include detection of the antigen based on immunological properties, physical properties, enzymatic properties and combinations thereof, or detection of a nucleic acid molecule encoding the antigen based on nucleic acid amplification.

Description: METHODS AND COMPOSITIONS FOR SCREENING FOR OR MODULATING A TUMOR ASSOCIATED ANTIGEN TECHNICAL FIELD The present invention is generally directed toward screening for or modulating a tumor associated antigen. The invention is more particularly related to detecting a complement Factor H-related protein, or a nucleic acid molecule encoding such a protein, associated with the presence of cancer, and to modulating the presence or activity of such a protein. BACKGROUND OF THE INVENTION The detection of new tumors or the recurrence of tumors remains an unfulfilled goal of humankind, despite enormous expenditures of both financial and human resources over the last twenty-five plus years. A number of cancers are treatable if detected at an early stage, but unfortunately go undetected in many patients for lack of a reliable screening procedure. For illustrative purposes, background for a particular cancer, bladder cancer, is described in more detail and is representative of cancers in need of new approaches, which the invention disclosed herein provides. Bladder cancer is the fifth most common cancer in the United States. The American Cancer Society estimated that a total of 52,000 new cases would be detected in 1994 and that there would be 10,000 deaths resulting from this disease. Bladder cancer is more common in men than in women by a ratio of approximately three to one and has been shown to be highly associated with smoking as well as exposure to certain dyes. Carcinoma of the urinary bladder is the fourth most common malignancy among American men, and the eighth among women. Transitional cell carcinoma (TCC) is the most common type of bladder cancer representing greater than 90% of all cases. The remaining cases are squamous cell carcinomas (7%), adenocarcinomas (2%), and undifferentiated carcinomas (1%). The diagnosis and management of TCC is often performed as follows. The patient presenting with such symptoms as hematuria or dysuria in the absence of infection undergoes a cystoscopy at which time the tumor is visualized. Although this procedure is invasive and unpleasant, it is highly accurate in predicting malignancy and is, thus, considered the gold standard. Urine cytology (i.e., the identification of tumor cells in voided urine) is also performed, and the combined results of the two methods may lead to an increase in sensitivity over that of cystoscopy alone. This is due to the fact that cytology occasionally allows detection of tumors which are not visible during cystoscopy, for example, flat tumors of the bladder (TIS) or those in the upper end of the bladder or the upper urinary tract. Transurethral biopsy and resection are then usually performed with this procedure removing the apparent lesion as well as providing information as to the grade and stage of the tumor. The tumor is typically graded from GO to G4 in decreasing state of differentiation. As with most cancers, the less differentiated the tumor the more aggressive the disease. With respect to stage or extent of invasion, TCC's of the bladder may be classified as superficial papillary (Ta and Tl), muscle invasive (T2 and greater), or the relatively uncommon tumor in situ (TIS). The extent of invasion dictates the type of therapeutic approach employed and the follow-up procedures to monitor for disease recurrence. Individuals with invasive TCC (Stage T2, T3, and T4) typically have poor prognoses. They are usually treated by radical cystectomy; however, in some cases the patient is unable to tolerate this surgery and is treated by radiation therapy or chemotherapy instead. This latter subgroup is monitored for disease recurrence by cystoscopy and urine cytology. Approximately 75% of TCC patients are initially diagnosed as having either Ta or Tl disease. In part because bladder cancer is multifocal, initial resection and treatment of these patients is curative in less than half of the cases. Although patients presenting with Ta TCC usually recur, their tumors tend to be low grade, and only 10-15% of the tumors will progress to muscle invasive disease. In contrast, Tl patients will progress 30-50% of the time. Superficial TCC is usually treated by transurethral resection, intravesical therapy, or fulguration, and follow-up is usually by cystoscopy and voided urine cytology. As mentioned above, current practice includes a preliminary diagnosis of TCC by cystoscopy and urine cytology, confirmatory diagnosis and staging and grading by biopsy, and routine follow-up of superficial and some invasive TCC by cystoscopy and urine cytology. Recurrence, especially within the first 12 months, is common, even when tumors have been diagnosed and treated prior to invasion of the bladder muscle. Therefore, patients with superficial TCC are typically monitored every three months for the first two years and, if there is no recurrence, every six months during the following year. Because cystoscopy is invasive and unpleasant and because urine cytology, although highly specific, is of variable reliability in detecting recurrence, there is a significant need for alternative diagnostic approaches. Accordingly, there is a need in the art for a non-invasive diagnostic method with reliability in detecting occurrence or recurrence. The present invention fulfills this need and further provides other related advantages. SUMMARY OF THE INVENTION Briefly stated, the present invention provides a variety of methods and compositions for screening for cancer, and for treating tumor cells. The screening methods and compositions may be used on a one-time basis when cancer is suspected or on a periodic basis, e.g., to monitor an individual with an elevated risk of acquiring or reacquiring cancer. In one aspect, the present invention provides a method of screening for a cancer comprising the step of detecting the presence of a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, the nucleic acid molecule characterized by the ability of the nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO: 19, respectively). Preferred embodiments of the methods for either aspect include detection of the antigen based on immunological properties, physical properties, enzymatic properties or combinations thereof, and detection of a nucleic acid molecule encoding the antigen by amplification of the molecule. In a related aspect, the present invention provides a method of treating a tumor cell comprising the step of modulating a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, the nucleic acid molecule characterized by the ability of the nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO:17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO: 19, respectively). In another aspect, the present invention provides agents that modulate a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen. In one embodiment, an agent is provided that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, the nucleic acid molecule characterized by the ability of the nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO:19, respectively), for use as a medicament to treat a tumor cell. In another embodiment, a composition is provided comprising an agent that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, the nucleic acid molecule characterized by the ability of the nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO:17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO: 19, respectively), in combination with a pharmaceutically acceptable carrier or diluent. In another embodiment, the present invention provides for use of an agent that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, the nucleic acid molecule characterized by the ability of the nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO: 18 and SEQ ID NO: 19, respectively), for the manufacture of a medicament for the treatment of a tumor cell. These and other aspects of the present invention will become evident upon reference to the following detailed description and attached drawings. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows the gel electrophoresis of the first-step RT-PCR amplification products, with lanes 1 to 10 beginning at the right side of the gel as lane 1. Lane 1 : X44.1 mRNA; Lane 2: HTB-5 mRNA; Lane 3: HTB-9 mRNA; Lanes 4, 5, 6 same targets as 1, 2, 3 respectively except the RT primer was OligodT 16 . Lanes 7 and 10, DNA molecular weight markers at 2000, 1500, 1000, 700, 500, 400, 300, 200, 100, and 50 base pairs. Lanes 8 and 9 are PAW 109, the kit positive control, at the expected size of 311 base pairs. Figure 2 shows the gel electrophoresis of the second-step PCR amplification products, with lanes 1 to 10 beginning at the right side of the gel as lane 1. Lane 1 : X44.1 product (reaction 1, lane 1) with primers 753RT and 412M; Lane 2, HTB-5 product (reaction 1, lane 2) with 753RT and 412M; Lane 3, HTB-9 product (reaction 1, lane 3) with 753RT and 412M. Lane 4, PAW109 (reaction 1, lane 8) product with 753RT and 42M primers; Lane 5, X44.1 product with 753RT and 42M primers; Lane 6, HTB-5 product with 753RT and 42M primers; Lane 8, HTB-9 product with 753RT and 42M primers. Lanes 7 and 11, DNA molecular weight markers. Lane 9, PAW 109 product (reaction 1, lane 8) with DM152 and DM151 primers; Lane 10, PAW109 product (reaction 1 , lane 8) with 753RT and 412M primers. Figure 3 shows stimulation by MAb X52.1 of the complement-mediated lysis of rabbit red blood cells. The extent of lysis is shown after 45 and 1 17 minutes with complement alone and in the presence of X52.1 at concentrations of 10 nM and 30 nM. Figure 4 shows stimulation by MAb X52.1 of the complement-mediated lysis of HL-60 human myeloid cells. The extent of lysis is shown after 120 minutes (a) with complement alone, (b) in the presence of X52.1 at a concentration of 10 nM, and (c) in the absence of complement. Figure 5 shows the gel electrophoresis of amplification products resulting from RT-PCR performed with three primer sets derived from human complement Factor H (lanes 1 to 10 beginning at the left side of the gel with the left side set of numbers 1-4 on the Figure representing lanes 1-4, the middle set of numbers 1-4 representing lanes 6-9 with lane 5 preceding, and the right side set of numbers 1-4 representing lanes 11-14 with lane 10 preceding). Lane 1 : HTB-9 product with primers 1040RT and 42M; Lane 2: HeLaS3 product with primers 1040RT and 42M; Lane 3: NHEK product with primers 1040RT and 42M; Lane 4: LS174T product with primers 1040RT and 42M; Lane 6: HTB-9 product with primers 1040RT and 410M; Lane 7: HeLaS3 product with primers 1040RT and 410M; Lane 8: NHEK product with primers 1040RT and 410M; Lane 9: LS174T product with primers 1040RT and 410M; Lane 1 1 : HTB-9 product with primers 3610RT and 2910M; Lane 12: HeLaS3 product with primers 361 ORT and 2910M; Lane 13: NHEK product with primers 361 ORT and 2910M; Lane 14: LS174T product with primers 3610RT and 2910M; Lanes 5 and 10: DNA molecular weight markers. Figure 6A shows a partial DNA sequence from clone pRBB9FH410 (SEQ ID NO:22) and Figure 6B the corresponding deduced amino acid sequence (SEQ ID NO: 24), as compared to the DNA and amino acid sequences for human CFH (SEQ ID NOS: 21 and 23 respectively). Figure 7A shows three partial DNA sequences from clone pRBS3FH2910 (SEQ ID NOS: 26-28) and Figure 7B the corresponding deduced amino acid sequences (SEQ ID NOS: 30-32), as compared to the DNA and amino acid sequences for human CFH (SEQ ID NOS: 25 and 29, respectively). Figure 8A shows two partial DNA sequences from clone pZS3FH2576 (SEQ ID NOS: 34 and 35) and Figure 8B the corresponding deduced amino acid sequences (SEQ ID NOS: 37 and 38), as compared to the DNA and amino acid sequences for human CFH (SEQ ID NOS: 33 and 36, respectively). DETAILED DESCRIPTION OF THE INVENTION As noted above, the present invention is directed, in one aspect, toward methods of screening for cancer. As disclosed in the present invention, a protein antigen has been found to be associated with the presence of cancer ("tumor- associated") and found to survive in detectable concentrations in samples from warm- blooded animals, such as humans. The present disclosure describes, for example, the purification of a tumor-associated antigen from cancer patients, the generation of antibodies to the antigen, the characterization of the antigen by physical and biological properties, the development of immunoassays and non-immunoassays for the detection of the antigen or a nucleic acid molecule encoding the antigen, the evaluation of samples from normal individuals and cancer patients, demonstration of the production of the antigen by cancer cells, the determination that the antigen corresponds to protein products related to human complement Factor H, and the inhibition of biological activity of the antigen. A wide variety of cancers may be screened. Representative examples of such cancers include urogenital, renal, head/neck and lung. Urogenital cancers include bladder, cervical and prostate. Head/neck cancers include cancers of the oral cavity, mouth and esophagus. As used herein, the term "screening for" includes detecting, monitoring or diagnosing. It will be evident to those in the art that if one wishes to screen for a particular type of cancer, this choice will guide the selection of a particular source of cell, tissue or sample to be tested. A sample in general may be a liquid or solid (e.g., cellular) sample taken from a tissue or organ, or after having been in contact with a tissue or organ. For example, a prostate sample includes a sample taken from a prostate or after having been in contact with a prostate. Representative types of prostate samples include prostate scraping and prostate tissue biopsy. A head/neck sample includes a sample taken from a head/neck or after having been in contact with a head/neck. Representative types of head/neck samples include swabs, scrapings and tissue biopsy of the oral cavity and esophagus. A lung sample includes a sample taken from a lung or after having been in contact with a lung. Representative types of lung samples include bronchial wash, sputum and tissue biopsy of the lung. A bladder sample includes a sample taken from a bladder or after having been in contact with a bladder. Representative types of bladder samples include urine, bladder wash, bladder scraping and bladder tissue biopsy. Urine may be voided or pre- voided (i.e., in a bladder). Urine may be removed from a bladder by using, for example, a catheter or a needle. A cervical sample includes a sample taken from a cervix or after having been in contact with a cervix. Representative types of cervical samples include cervical swab, cervical wash, cervical scraping and cervical tissue biopsy. Pretreatment of a sample may be desirable. For example, in the case of urine samples neutralizing the pH with buffer may be desirable. The detection, isolation, characterization and identification of a protein antigen present in specimens derived from patients with cancer, but absent in specimens from normal individuals, indicates that this antigen is either a product of the cancer cells or is for some other reason present in specimens from these patients. If the antigen is expressed by cancer cells, it may be present in the supematants taken from cultured human cancer cell lines at levels adequate to be measured by enzyme immunoassay specific for the antigen. cDNA derived by reverse transcriptase-polymerase chain reaction (RT-PCR) amplification from mRNA isolated from the same cancer cells can be used as well to provide evidence for expression of the gene which encodes for a product which is identical or very similar to the identified antigen. As disclosed herein, both types of experimental approaches confirm the expression of the antigen by cancer cell lines (e.g., bladder, cervical, renal and prostate cancer cell lines). The tumor-associated protein antigen of the present invention has been determined, by sequence comparisons, suφrisingly to be human complement Factor H- related. As cancer cells may produce more than one form of the protein, as used herein the term "human complement Factor H-related" refers to the human complement Factor H protein and variants thereof. The variants may be the result of mutations, alternate splicing or recombination events that alter nucleic acid molecules encoding human complement Factor H. In general, the amino acid sequence identity between a human complement Factor H-related protein from a tumor cell and human complement Factor H will be at least about 50%. More typically, the amino acid sequence identity will be at least about any integer from (and including) 50% to 100%, such as at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95% identity. Variants that are nearly identical to human complement Factor H have at least about 85% or 90% identity. As used herein, amino acid sequence "identity" is determined by the alignment of amino acid sequences and establishment of identical amino acid residues using the program GeneJockey II (1993) for Macintosh (Philip L. Taylor, published by Biosoft, Cambridge, UK). The program is run in the amino acid homology mode, using program default parameters. In the comparison of two sequences aligned by the program, the percent identity is calculated only for those positions where there is an amino acid residue present in both of the two sequences. In addition, a nucleic acid molecule encoding for a human complement Factor H-related protein will typically hybridize under moderately stringent conditions to one or the other or both of two primer pairs (42M/1040RT or 2910M/3610RT), as described below. This reflects conservation of certain sequences (disclosed herein) for tumor-associated human complement Factor H-related antigen. A protein may generally be identified as a tumor-associated human complement Factor H-related antigen based on the ability of a nucleic acid molecule encoding the protein to hybridize under moderately stringent conditions to one or the other or both of two primer pairs (42M/1040RT or 2910M/3610RT), as described below. Based on the disclosure herein, in combination with the methodologies known in the art, it will be evident to those in the art whether a protein is a tumor-associated human complement Factor H-related antigen, or whether a nucleic acid molecule encodes such a protein. The antigen may be isolated in substantially pure form. Briefly, for example, urine samples of bladder cancer patients are clarified (e.g., by centrifugation) and concentrated (e.g., by hollow fiber concentrator). The concentrated sample is chromatographed on heparin agarose, and bound material eluted using a linear buffered NaCI gradient. Pooled fractions are concentrated. Purity can be assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis ("SDS-PAGE") with appropriate protein stains. Alternatively, the antigen may be purified using an antibody against the antigen, as described for example below. Following isolation of antigen, the polypeptide constituents may be identified. Typically, polypeptides are resolved by separation (e.g., by gel electrophoresis) under denaturing conditions (e.g., sodium dodecyl sulfate). Approximate molecular weights of polypeptides are assigned by comparison of their mobility to the mobility of polypeptides of known molecular weights on SDS-PAGE. Isolated antigen yields from certain cancers, for example, a band with an apparent molecular weight of approximately 151 ,000 on SDS-PAGE under reducing conditions (i.e., in the presence of DTT which is 1,4-dithiothreitol). Rather unusually, this band exhibits a lower apparent molecular weight (of approximately 138,000) on SDS-PAGE under non-reducing conditions (i.e., in the absence of a reducing agent). This somewhat anomalous electrophoretic behavior provides a convenient means for identifying the antigen. Purified antigen, partially purified antigen or biological samples containing antigen may be used to produce antibodies that specifically bind to the antigen. Antibodies that specifically bind are those with an affinity of about 10 6 liters/mol or greater. Either polyclonal antibodies or monoclonal antibodies may be generated. Polyclonal antibodies may be produced by immunization of an animal and subsequent collection of its sera. It is generally preferred to follow the initial immunization with one or more booster immunizations prior to sera collection. Monoclonal antibodies are generally produced by the method of Kohler and Milstein (Nature 256:495-497, 1975; Eur. J. Immunol. 5:51 1-519, 1976). Briefly, the lymph nodes and/or spleens of an animal injected with antigen in pure or impure form are fused with myeloma cells to form hybrid cell lines ("hybridomas" or "clones"). Each hybridoma secretes a single type of immunoglobulin specific for the antigen and, like the myeloma cells, has the potential for indefinite cell division. Antigen in pure or impure form ("immunogen") is used for the immunization. Preferably, the animals are immunized with at least 100 ng each of the immunogen, most preferably greater than 500 ng each. For immunization, the immunogen may be adsorbed to a solid phase matrix, preferably to nitrocellulose paper. The paper is then introduced into the animal. Techniques for introduction of the adsorbed antigen preparation include implantation (U.S. Patent No. 4,689,220) or solubilization of the solid phase and injection of the solubilized material (Knudsen, Anal. Biochem. 747:285-288, 1985). The solid phase matrix may be solubilized in an appropriate organic solvent (e.g., DMSO) and either mixed with adjuvant or saline, or injected directly. Alternatively, the immunogen may be injected in the absence of a solid matrix and/or adjuvant. Injection or implantation may be intraperitoneal, intra-foot pad, subcutaneous, intramuscular or intravenous, but preferably intraperitoneal. The animals may also be injected with antigen complexed with adjuvant, such as Freund's adjuvant. Single or multiple booster immunizations are used. Between one and seven days prior to the fusion date, preferably on days one through four, intravenous injections of the immunogen may be given daily. Between one and seven days, preferably four days, after the administration of the final booster immunization, spleens or portions thereof are harvested from the immunized animals. At this time, the lymph nodes may also be harvested and included in the cell preparation. The harvested organs are minced using techniques which disrupt the structure of the organ, but which are not detrimental to the lymphocytes. The organs are preferably minced with scissors, passed through a mesh screen and mixed with growth medium to enrich the preparation for lymphocytes. The minced and strained tissue is harvested by centrifugation, then mixed with growth medium to form a cell suspension. The red blood cells may be lysed by adding a hypotonic or hypertonic solution to the cell suspension. A preferred method for cell lysis is to add distilled water to the suspensions and quickly return the suspensions to an isotonic state with a hypertonic sodium chloride solution. Any remaining tissue may be removed by filtration through gauze. The harvested cell suspension is then mixed with a myeloma cell line, preferably one which is syngeneic with the immunized animal. Myeloma cell lines from various species are widely available through, for example, American Type Culture Collection (ATCC), Rockville, Maryland. Myeloma cell lines commonly used include P3X63Ag8 (ATCC TIB 9), SP2/0-Agl4 (ATCC CRL 1581), FO (ATCC CRL 1646) and 210-RCY-Agl (Galfre et al., Nature 277:131, 1979). The myeloma cells are cultured in an appropriate mammalian cell growth medium, a variety of which are generally known in the art and available from commercial sources. Mammalian cell lines are routinely grown between 36°C and 40°C under conditions which maintain an optimal pH between 6.0 and 8.0, preferably about pH 7.2. pH may be maintained through the use of a variety of buffer systems known in the art. A preferred buffer system involves growing the cells in a bicarbonate buffer in a humidified incubator containing CO2, preferably about 7% CO2. The fusion between the lymphocytes from the immunized animal and the myeloma cells may be carried out by a variety of methods described in the literature. These methods include the use of polyethylene glycol (PEG) (Brown et al., J. Biol. Chem. 255:4980-4983, 1980) and electrofusion (Zimmerman and Vienken, J. Membrane Biol. (57:165-182, 1982). An electrofusion generator is commercially available from Biotechnologies and Experimental Research, Inc., San Diego, California. Following the fusion, the cells are plated into multi-well culture plates, preferably 96-well plates. A reagent which selectively allows for the growth of the fused myeloma cells over the unfused cells is added to the culture medium. A preferred selection technique uses HAT (hypoxanthine, aminopterin, thymidine) selection. Other selection techniques may also be used depending on the myeloma cell line chosen. Alternative methods of producing monoclonal antibodies utilize in vitro immunization techniques. Lymphocytes may be harvested from lymphoid organs, such as spleen or lymph nodes, or from whole blood as peripheral blood lymphocytes. The lymphocytes are put into culture in the presence of the appropriate immunogen. Often immunostimulatory polypeptides will be added to the culture medium concurrently. At various times following the culturing of the lymphocytes in vitro, the lymphocytes are harvested and fused with a myeloma cell line as described above. Other techniques for producing and maintaining antibody secreting lymphocyte cell lines in culture include viral transfection of the lymphocyte to produce a transformed cell line which will continue to grow in culture. Epstein-Barr virus (EBV) has been used for this technique. EBV transformed cells do not require fusion with a myeloma cell to allow continued growth in culture. Thymocytes may be used as a feeder layer to condition the medium for the fused cells. Alternatively, peritoneal macrophages or non-immune spleen cells may be used as a feeder layer. Another alternative is to use conditioned medium from thymocytes or macrophages. Thymocytes may be prepared from juvenile mice less than 8 weeks old. The thymus glands are harvested and minced using techniques which disrupt the thymus gland but are not detrimental to the thymocytes. This procedure is preferably carried out using scissors to mince the tissue, followed by passage of the tissue through a mesh screen. The minced and strained cell material is then harvested by centrifugation. Cell suspensions are made using growth medium. Any remaining connective tissue may be removed by filtration through gauze. At an appropriate time following the day the cells are fused, the fused cells (hybridomas) are then analyzed for the production of antibody against the antigen. This "screening" can be done by a wide variety of techniques, including Western blot, ELISA, immunoprecipitation, effect on biological activity assays and immunocytochemical staining. These techniques and others are well described in the literature. (See, for example, J. G. R. Hurrell (ed.), Monoclonal Hybridoma Antibodies: Techniques and Applications, CRC Press Inc., Boca Raton, Fla., 1982.) Introduction of a screening procedure permits further definition of antibodies of useful reactivity. For example, antigen purified from a biological sample of a patient with a bladder cancer may be used in any of the above-named techniques to define antibodies which react, for example, to determinants which are common to patients with the disease. Hybridomas which secrete antibodies of interest are maintained in culture. The cells are expanded in culture and at the same time may be cloned in such a manner as to obtain colonies originating from single cells. This provides for the monoclonal nature of the antibodies obtained from the hybridomas. A wide variety of techniques exist for cloning cells, including limiting dilution, soft agar cloning and fluorescence-activated cell sorting. Once clones of cells are obtained, they are re-assayed for the production of the antibody of interest. These cells are then expanded in culture to allow for the production of larger amounts of the antibody. Methods for expansion of the cells include maintaining the cells in culture, placement of the cells in a bioreactor or other type of large-scale cell culture environment, or culturing the cells using various agar or gelatin carrier matrices. Antibodies are then isolated from the cell culture media. Antibodies may be purified from conditioned media or ascites fluid by a variety of methods known in the art. These methods include ammonium sulfate precipitation, ion exchange chromatography (see Hurrell, ibid.) and high pressure liquid chromatography using a hydroxylapatite support (Stanker et al., J. Immunol. Methods 76:157, 1985). A preferred method for purifying antibodies from conditioned media or ascites fluid utilizes a commercially available Protein A-Sepharose ® CL-4B column or Protein G Sepharose ® (Pharmacia, Piscataway, NJ; Sigma, St. Louis, MO) or ABX mixed ion exchange resin (JT Baker, Phillipsburg, NJ). Antibodies may be purified with these columns using conditions suggested by the manufacturer. As disclosed herein, the antigen which is found to be associated with the presence of cancer may be detected in a wide variety of ways, including by detecting the antigen itself or a nucleic acid molecule encoding the antigen. Methods for detecting the presence (i.e., qualitative or quantitative) of the antigen include those based on its physical properties, immunological properties, enzymatic properties and combinations thereof. For example, regarding physical properties, the antigen's unique polypeptide mobility on SDS-PAGE under reducing and non-reducing conditions may be exploited for a determination as to whether antigen is present in a sample. More specifically, for example, as described herein, a polypeptide with an apparent molecular weight on SDS-PAGE of about 151,000 under reducing conditions exhibits a lower molecular weight of about 138,000 under non-reducing conditions. Alternatively, the presence of antigen may be detected by immunological means. The means for detecting the presence of antigen may be in a direct or indirect test format. In a direct test format, that which is observed or measured is proportional to (i.e., directly reflective of) antigen present in a sample. Conversely, in an indirect test format, that which is observed or measured is inversely proportional to (i.e., indirectly reflective of) antigen present in a sample. Indirect formats include competitive and inhibition assay formats. As used herein, the term "antibody" includes both polyclonal and monoclonal antibodies; and may be an intact molecule, a fragment thereof, or a functional equivalent thereof; and may be genetically engineered. Examples of antibody fragments include F(ab')2, Fab', Fab and Fv. Detection may be, for example, by Western blot analysis utilizing antigen immobilized on nitrocellulose or Immobilon or similar matrix, in conjunction with specific antibodies to the antigen. Detection can also be achieved by immunoassay. In one embodiment, antigen is isolated from a sample and contacted with an appropriate detection antibody. Antigen may be isolated by capture on a solid support (e.g., heparin agarose) or with a "capture" antibody prior to or simultaneous with a "detection" antibody. In another embodiment, immunocomplexes are formed between an antibody and antigen, without prior purification of the antigen. Incubation of a sample with an antibody is under conditions and for a time sufficient to allow immunocomplexes to form. Detection of antigen by immunological means is also amenable to quantification where it is desired to determine the amount of antigen. Detection of one or more immunocomplexes formed between antigen and an antibody specific for the antigen may be accomplished by a variety of known techniques, including radioimmunoassays (RIA) and enzyme linked immunosorbent assays (ELISA). The immunoassays known in the art include the double monoclonal antibody sandwich immunoassay technique of David et al. (U.S. Patent 4,376,110); monoclonal-polyclonal antibody sandwich assays (Wide et al., in Kirkham and Hunter (eds.), Radioimmunoassay Methods, E. and S. Livingstone, Edinburgh, 1970); the "western blot" method of Gordon et al. (U.S. Patent 4,452,901); immunoprecipitation of labeled ligand (Brown et al., J. Biol. Chem. 255:4980-4983, 1980); enzyme-linked immunosorbant assays as described by, for example, Raines and Ross (J. Biol. Chem. 257:5154-5160, 1982); immunocytochemical techniques, including the use of fluorochromes (Brooks et al., Clin. Exp. Immunol. 39: 477, 1980); and neutralization of activity (Bowen-Pope et al., Proc. Natl. Acad. Sci. USA 57:2396-2400, 1984). In addition to the immunoassays described above, a number of other immunoassays are available, including those described in U.S. Patent Nos.: 3,817,827; 3,850,752; 3,901,654; 3,935,074; 3,984,533; 3,996,345; 4,034,074; and 4,098,876. For detection purposes, the antibodies may either be labeled or unlabeled. When unlabeled, the antibodies find use in agglutination assays. In addition, unlabeled antibodies can be used in combination with other labeled antibodies (second antibodies) that are reactive with the antibody, such as antibodies specific for immunoglobulin. Alternatively, the antibodies can be directly labeled. Where they are labeled, the reporter group can include radioisotopes, fluorophores, enzymes, luminescers, or visible particles (e.g., colloidal gold and dye particles). These and other labels are well known in the art and are described, for example, in the following U.S. patents: 3,766,162; 3,791,932; 3,817,837; 3,996,345; and 4,233,402. Typically in an ELISA assay the target antigen (for a competitive or inhibition assay format) or immobilized capture antibody is adsorbed to the surface of a microtiter well. Residual protein-binding sites on the surface are then blocked with an appropriate agent, such as bovine serum albumin (BSA), heat-inactivated normal goat serum (NGS), or BLOTTO (buffered solution of nonfat dry milk which also contains a preservative, salts, and an antifoaming agent). The well is then incubated with a sample suspected of containing antigen. The sample can be applied neat, or, more often, it can be diluted, usually in a buffered solution which contains a small amount (0. l%-5.0% by weight) of protein, such as BSA, NGS, or BLOTTO. After incubating for a sufficient length of time to allow specific binding to occur, the well is washed to remove unbound protein and then incubated with a detection antibody labeled with a reporter group, or an anti-immunoglobulin antibody labeled with a reporter group. The reporter group can be chosen from a variety of enzymes, including horseradish peroxidase, beta- galactosidase, alkaline phosphatase, and glucose oxidase. Sufficient time is allowed for specific binding to occur, the well is again washed to remove unbound conjugate, and the substrate for the enzyme is added. Color is allowed to develop and the optical density of the contents of the well is determined visually or instrumentally. In one preferred embodiment of the present invention, a reporter group is bound to the detection antibody. The step of detecting an immunocomplex involves removing substantially any unbound antibody and then detecting the presence or absence of the reporter group. In another preferred embodiment, a reporter group is bound to a second antibody capable of binding to the antibody specific for antigen. The step of detecting an immunocomplex involves (a) removing substantially any unbound antibody, (b) adding the second antibody, (c) removing substantially any unbound second antibody and then (d) detecting the presence or absence of the reporter group. Where the antibody specific for the fragment is derived from a mouse, the second antibody is an anti-murine antibody. In a third preferred embodiment for detecting an immunocomplex, a reporter group is bound to a molecule capable of binding to the immunocomplex. The step of detecting involves (a) adding the molecule, (b) removing substantially any unbound molecule, and then (c) detecting the presence or absence of the reporter group. An example of a molecule capable of binding to the immunocomplex is protein A. It will be evident to one skilled in the art that a variety of methods for detecting the immunocomplex may be employed within the present invention. Reporter groups suitable for use in any of the methods include radioisotopes, fluorophores, enzymes, luminescers, and visible particles (e.g., colloidal gold and dye particles). As disclosed herein, this antigen, which is associated with the presence of cancer, is bound by complement factor fragment C3b. Therefore, C3b may be used in assays (such as those described above) that utilize a capture molecule and a detection molecule for detecting antigen. For example, C3b may be immobilized on a solid support and used to capture antigen when contacted with a sample containing antigen. Another molecule which is specific for antigen, such as an antibody, may be used to detect any antigen bound to immobilized C3b. It may be desirable to wash the immobilized C3b, after introducing a sample suspected of containing antigen, prior to and/or subsequent to contacting with a detection molecule. Alternatively, this antigen possesses enzyme cofactor activity, e.g., whereby it acts as a cofactor for the digestion of C3b by Factor I of the complement system. Therefore, the presence of antigen may be determined by contacting a sample (suspected of containing antigen) with C3b and Factor I, and assaying for the digestion of C3b. In the presence of antigen and Factor I, the C3b α' fragment at a molecular weight of about 108,000 disappears with the concurrent appearance of fragments with molecular weights of 67,000 and 47,000. This digestion may be detected by a variety of ways, including SDS-PAGE. Alternatively, rather than detecting the antigen itself, a nucleic acid molecule encoding the antigen can be detected. Such a nucleic acid molecule may be a deoxyribonucleic acid (DNA) or a ribonucleic acid (RNA). Generally, a nucleic acid molecule encoding for the antigen is detected by amplification of the nucleic acid. A variety of methods may be utilized in order to amplify a selected sequence, including, for example, RNA amplification (see Lizardi et al., Bio/Technology 6:1197-1202, 1988; Kramer et al., Nature 359:401-402, 1989; Lomeli et al., Clinical Chem. 35(9): 1826- 1831, 1989; U.S. Patent No. 4,786,600), and DNA amplification utilizing ligase chain reaction ("LCR") or polymerase chain reaction ("PCR") (see U.S. Patent Nos. 4,683,195, 4,683,202, and 4,800,159) (see also U.S. Patent Nos. 4,876,187 and 5,01 1,769, which describe an alternative detection/amplification system comprising the use of scissile linkages), or other nucleic acid amplification procedures that are well within the level of ordinary skill in the art. With respect to PCR, for example, the method may be modified as known in the art. Transcriptional enhancement of PCR may be accomplished by incorporation of bacteriophage T7 RNA polymerase promoter sequences in one of the primary oligonucleotides, and immunoenzymatic detection of the products from the enhanced emitter may be effected using anti-RNA:DNA antibodies (Blais, Appl. Environ. Microbiol. 60:348-352, 1994). PCR may also be used in combination with reverse dot-blot hybridization (Iida et al., FEMS Microbiol. Lett. 774:167-172, 1993). PCR products may be quantitatively analyzed by incoφoration of dUTP (Duplaa et al., Anal. Biochem. 272:229-236, 1993), and samples may be filter sampled for PCR-gene probe detection (Bej et al., Appl. Environ. Microbiol. 57:3529-3534, 1991). Primers for the amplification of a selected sequence should be selected from sequences that are highly specific to the antigen and form stable duplexes with the target sequence. The primers should also be non-complementary, especially at the 3' end, should not form dimers with themselves or other primers, and should not form secondary structures or duplexes with other regions of DNA. In general, primers (such as those described in greater detail below) of about 18 to 30 nucleotides are preferred, and can be easily synthesized using techniques well known in the art. PCR products, and other nucleic acid amplification products, may be quantitated using techniques known in the art (Duplaa et al., Anal. Biochem. 272:229-236, 1993; Higuchi et al., Bio/Technology 77:1026-1030). A preferred embodiment involves assaying for the presence of specific messenger RNA (mRNA) encoding the antigen. More specifically, for example, as described herein, a cell sample may be lysed and the mRNA isolated, amplified and examined for the presence of mRNA specific for the antigen. A variety of procedures may be used to detect the presence of antigen-specific mRNA. A particularly preferred method includes RT-PCR (Reverse Transcriptase based Polymerase Chain Reaction) amplification of mRNA. Detecting the presence of antigen in a cell, tissue or sample has a variety of uses. For example, the present invention may be used for diagnostic puφoses to screen warm-blooded animals, such as humans, for cancer (or a particular cancer depending upon the source of the particular cell, tissue or sample). Similarly, the present invention may be used to monitor warm-blooded animals. In particular, a preferred use is to follow patients who have been previously diagnosed and treated for cancer. Patients who are in remission (or may in fact be cured) can be monitored for the reappearance of cancer. It will be evident to those in the art that it may be desirable to use the present invention in conjunction with one or more other tests for cancer (or a particular cancer) to confirm positive or negative results obtained from use of the present invention. The unexpected presence of a complement Factor H-related protein in cell culture supematants from epithelial cancer cells (Example VI, Table 7), and the demonstration that its mRNA is produced by cancer cells (Example VI, Table 7), suggest that it plays a significant role in cancer biology. Data presented below (Example III.F) demonstrate that a biological activity of the antigen is to accelerate the complement Factor I-mediated degradation of C3b. The role of C3b in vivo is the assembly of the membrane attack complex (MAC) prior to lysis of an appropriate target. Because these proteins are members of the Alternative Complement Pathway, cell lysis may take place independent of the presence of circulating antibodies to any of the cancer cell antigens. Although not wishing to be bound by theory, in view of the activity of the antigen described herein, its production by cancer cells may locally promote the degradation of C3b, thereby inhibiting the formation of the MAC and preventing tumor cell lysis by complement. Since the production of the antigen by tumor cells may afford a survival advantage, interrupting the production of the antigen or blocking its decay accelerator activity restores susceptibility of the tumor to complement-mediated cell lysis, thus offering a new approach to cancer therapy. Irrespective of the exact function(s) of the complement Factor H-related protein in tumor biology, the present invention provides for the modulation of the antigen as a means for treating cancers. It will be evident to those of ordinary skill in the art that the antigen may be modulated in a variety of ways. A preferred method of interrupting the production of the antigen is by use of DNA, or PNA (peptide nucleic acid), constructs with base sequence complementary to the antigen's mRNA. Such an approach is generically termed antisense technology. Typically, the complement Factor H antisense DNA is inserted into an appropriate vector (virus) which delivers it to the tumor cells. Once inside the target cells, the antisense construct would specifically bind to mRNA coding for the complement Factor H-related protein, thereby preventing its translation. Primary among the other methods which could be used to interrupt production of the antigen would be the use of specific molecules which block the transcription of the specific gene or genes coding for the complement Factor H- related protein. Chemicals designed to block the ability of the tumor cell to produce antigen would preferably be delivered in the vicinity of the tumor, rather than systemically, since systemic introduction of such materials could decrease the normal production of complement Factor H by the liver, compromising the host's ability to regulate complement activity. In modulation of antigen production, it is desired to eliminate the production of all complement Factor H-related protein by tumor cells. Another approach to antigen modulation is to use reagents to inhibit the activity of complement Factor H activity. Unlike inhibition of antigen production, the dosage used with these reagents should preferably result in an inhibition of 70%-95% of the Factor H activity. One family of such inhibitors — monoclonal antibodies, or fragments which bind the antigen at a site which blocks its ability to degrade C3b — is presented as a representative example of modulation of antigen activity as an approach to cancer therapy (Example VII). In this example, certain antibodies which bind antigen are shown to accelerate the complement-mediated lysis of rabbit red blood cells and HL-60, a human tumor cell line. With these reagents, as with those described above, delivery should preferably be administered to the tumor site, rather than systemically. For the antibodies described above, reagent affinities should be at least about IO 6 liters/mole and doses should be within the range of about 0.01 μg/kg body weight to 10 mg/kg body weight. In addition, the preferred type of tumor to be treated in this manner would be distinctly separate from the circulatory system, since blood itself contains high concentrations of complement Factor H. An antibody may be replaced by, or supplemented with, any peptide or other organic molecule which specifically binds to complement Factor H-related protein and blocks its biological activity. The above-described molecules (antibodies, peptides, organic compounds and antisense nucleic acids or peptide nucleic acids) are representative examples of agents that may modulate a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding the antigen, for use as a medicament to treat a tumor cell. Such agents may be used for the manufacture of a medicament for the treatment of a tumor cell. Further, such agents may be combined with a pharmaceutically acceptable carrier or diluent to from a composition. Additional components, such as traditional chemotherapeutic compounds, may be included with such an agent or a composition thereof. The following examples are offered by way of illustration and not by way of limitation. EXAMPLES EXAMPLE I DEVELOPMENT OF MONOCLONAL ANTIBODIES A. Antigen The antigen source for immunization was a pool of Heparin-Agarose fractionated urines from clinically diagnosed bladder cancer patients. (The purification method is described in detail in Example III.A.l. below.) Twenty-four hour urine samples were centrifuged in a Beckman centrifuge (Fullerton, CA), Model #J2-21, S N 5539, using a JA-10 rotor at 6,000 φm for 20 minutes. The clarified urine sample was then concentrated using an Amicon stirred cell, 76 mm, (cat# 5124) fitted with a YM30 membrane MWCO 30,000 dalton (Amicon, cat# 13742) or a Microgon hollow fiber concentrator, 50,000 MWCO (cat# M15S-260-01N) to approximately 100X concentration. The concentrated sample was diluted 1 :1 with 25 mM Tris-HCl pH 7.4 and loaded onto a column of Heparin- Affigel (BioRad, Richmond, CA, cat# 153-6173), equilibrated in 25 mM Tris-HCl pH 7.4, at a flow rate of 2.0 mL/min. The sample was followed with equilibration buffer until the A280 elution profile returned to background. Bound material was eluted with a linear NaCI gradient from 0 to 250 mM NaCI in 25 mM Tris-HCl pH 7.4, followed by 50 mL of 250 mM NaCI, 25 mM Tris-HCl, pH 7.4, and finally 20 mL of a 10X PBS, pH 7.4, solution. Five mL fractions were collected and fractions from the trailing half of the elution peak were pooled. Pooled fractions were concentrated with an Amicon stirred cell, 43 mm (cat# 5122), fitted with a YM30 membrane, MWCO 30,000 dalton (cat# 13722). Fractions comprising the pooled antigen are shown below: Pool I Patient 1 fractions 13-31 Patient 1 fractions 14-30 Patient 2 fractions 11-19 Patient 3 fractions 11-24 Pool II (1.5 mg/mL) Pool I l mL Patient 2 fractions 1 1-19 1 mL Patient 3 fractions 1 1 -24 1 mL Patient 4 fractions 1 1 -20 1 mL B. Immunizations Five female BALB/c mice, of 8-10 weeks of age, were immunized intraperitoneally with 0.2 mL of a 1:1 emulsion of Pool II in Freund's Complete Adjuvant (Difco, Detroit, MI). Three weeks later, booster immunizations of 0.1 mL containing 10 μg of protein of an emulsion in incomplete Freund's Adjuvant was administered to the rear footpads and peritoneum. Ten days later each mouse was sampled for antibody response via retro-orbital bleeds and the sera were tested via an ELISA described below for titers. Mouse number 340 showed the highest titer and was chosen for fusion four days after boosting in the footpads and peritoneum with 15 μg of Pool II in phosphate buffered saline. C. Fusion Four days after the last immunization animal # 340 was sacrificed, the popliteal and inguinal lymph nodes and the spleen were collected and used for fusion. Fusion was carried out by a modification of the method of Fazekas De St. Groth and Scheidigger, J. Immunol. M. 35:1-21, 1980. The parent hybridoma line FO (ibid.), obtained from the ATCC, was used for fusion, at a ratio of one to five lymphocytes. PEG-DMSO (Sigma, St. Louis, MO) fusogen was used, and the cells plated out in Iscove's Modified Dulbecco's Medium (IMDM) with penicillin-streptomycin and hypoxanthine/thymidine (HT) supplement at a density of 2 x IO 4 cells/well with 2.58 x 10 3 peritoneal macrophages from unimmunized BALB/C mice added as feeders. The fusion was divided into two parts, in the first part forty-eight 96 well plates were seeded at the above density in media containing 1% fetal bovine serum (FBS). The second part consisted of 49 plates seeded at the same density in media containing 10% FBS. A total of 97 plates, or 9,312 wells were used. The plates were incubated at 37°C in 7% CO 2 at 100% humidity. The next day 100 μl of selective media consisting of IMDM-HT with 2x methotrexate (8 x 10 "7 M) and appropriate FBS concentration was added. The plates were returned to the incubator and not disturbed for six days. On day seven the plates were removed from the incubator and approximately 150 μl of media was removed via aspiration with a sterile eight place manifold. Complete IMDM with HT and appropriate FBS was added to each well using a Brinkman eight place pipette. The plates were returned to the incubator for another five to six days before screening. The fusion plates were examined each morning for wells showing growth levels suitable for screening, and were analyzed that day. Within one week of the fusion, the plates containing the 1% FBS medium were clearly lagging in growth, and were therefore supplemented to 10% FBS. Thereafter, those wells selected from the plates initially plated in 1% FBS were designated as MOFI-followed by a number indicating the order of selection, those from the 10% FBS plates were designated with the MOFX prefix. D. Post-Fusion Cell Culture Wells selected via the screening assays were immediately transferred to 24 well plates containing 1 mL of complete IMDM containing 10% FBS. A sample of cells was also used to immediately re-clone the hybridomas by a serial limiting dilution procedure. This consisted of transferring a 10 μl sample of cells from the chosen well of the 96 well plate to the first well of a fresh 96 well plate previously filled with 100 μl of complete IMDM with 10% of a cloning supplement prepared from murine macrophages and thymocytes (Condimed, Boehringer-Mannheim Coφ., Indianapolis, IN). Cells from the first well were serially diluted in the first column of wells by transferring 100 μl from the first well to the second, then from the second to the third, etc. The remaining 100 μl removed from the last well of the column is transferred back to the first well. The wells of the first column were then serially diluted across the plate by transfer of 50 μl of cell suspension using an 8 place pipette. Finally, 100 μl of cloning media was added to each well, and the plates incubated for approximately two weeks before subclones were ready for re-screening. Following growth in the 24 well plates, the clones were transferred to six well plates with 5-6 mL of culture media, the plates were incubated until near confluent growth was observed. A sample of the cells were removed for storage in a cryogenic freezer in 5% DMSO in FBS, and the remaining cells were transferred to a T-75 flask with 10 mL media for producing spent media for further testing. E. Stabilization of Subclones Subclones were again subjected to testing via ELISA (described below) incoφorating an additional urine from a patient diagnosed as TCC+. Typically all subclones of a given original-evaluated well showed similar binding patterns and levels. Those showing loss of antibody production in all subclones were discarded, while those displaying loss in any examined subclone were subjected to another subcloning. This was repeated until all subclones showed comparable levels of expression. Nomenclature for each level of subcloning consisted of appending to the clone designation a period followed by the number of the selected subclone. F. Assays The titer assay was carried out by coating Pool II (Example I. A., above) antigen adjusted to 4 μg/mL in 0.1 M carbonate buffer, pH 9.6, directly to polystyrene plates. Each well received 50 μl of coating solution and the plate was covered and incubated at 37°C for 2 hours, after which time it was washed twice with phosphate buffered saline (PBS) in a Denley strip well washer. The plate was blocked by the addition of 100 μl of a 1% gelatin hydrolysate, 2% sucrose solution in 50mM Tris-HCl, pH 7.5, at 37°C for 1 1/2 hours (all reagents from Sigma). Following blocking, the plate was again washed twice with PBS, then two-fold serial dilutions of mouse serum, starting at 1 :100, into 10% normal horse serum in PBS, were added row- wise to the plate at 50 μl per well. The plate was incubated at 37°C for I hour, washed 4 times in PBS, and 50 μl of affinity purified goat anti-mouse IgG- horseradish peroxidase (HRP) conjugate (Tago, Burlingame, CA) diluted 1 :5000 in 10% horse serum in PBS added to each well. This was allowed to incubate for 1 hour at 37°C. The plate was washed with PBS 4 times, and 50 μl of substrate (K-Blue, ELISA Technologies, Lexington, KY) was added and the plate allowed to develop for 10 minutes at room temperature before stopping the reaction via the addition of 100 μl of 2M phosphoric acid solution in water (Sigma). The optical density of the wells were read at 450 and at 410 nm in a BioTek EL31 1 plate reader. Readings which were off scale at 450 nm were calculated from the corresponding reading at 410 nm by the method of Madersbacher and Berger, J. Immunol. M. 735. 121-124, 1991. The fusion was screened for antibody production by use of the following fusion screen. Antibody binding was tested with: (a) two clinically diagnosed patient urines, stages T2III and T3III, (diluted 1:80), (b) two pools of normal human urines (diluted 1 :15), (c) human type IV collagen (diluted to 4 μg/mL), all dilutions in 25mM Tris-HCl, and (d) pooled human red blood cells (Gamma Biologicals, Houston, TX) diluted into PBS and coated onto poly-lysine coated plates. All plates were blocked by washing with PBS with 0.1% Tween-20, and by the dilution of the media samples 1:5 into complete IMDM containing 10% FBS. Supernatant fluid (70 μL) of the wells chosen for screening were transferred to a well of a 96 well plate. To each well, 280 μl of diluent was added, and 50 μl was distributed to the test plate wells. The remaining steps of the assay were as for the titer assay, with the exception that the conjugate used was human serum adsorbed goat anti-mouse IgG-HRP conjugate (Kirkegaard and Perry Labs (KPL), Gaithersburg, MD) diluted 1 :5000 in 10% normal goat serum in PBS for all except the RBC plates. For the latter, an alkaline phosphatase conjugate of a similar antibody was used (KPL, Gaithersburg, MD) followed by use of PNPP (p-nitrophenyl phosphate) substrate. Controls were used for each assay, negative control was fresh IMDM with 10% FBS, positive controls were monoclonal anti-human collagen (Sigma C1926), and monoclonal anti-hlgA (Al.1.2.4, Bard Diagnostic Sciences, Inc., Redmond, WA), both of which showed high binding to all test antigens except the red blood cells. Criteria for selection were high binding to cancer urine plates (OD>l), low binding to normal urines and other test antigens (OD<0.5). Others which showed high antibody levels in different patterns with respect to the test antigens were also selected for potential research uses. Subclones were screened by several assays. First, the fusion assay was again used then, following expansion in culture of selected subclones, an abbreviated ELISA was employed using normal urine pool I and the two advanced stage urines used in the fusion assay. The testing was carried out at dilutions of 1 : 10 and 1 :100 for the early subclones, and an additional dilution of 1 :1000 for the later subclones. In several of the subclone assays the addition of urine from a patient with a lower grade cancer was included. From the 9,312 wells plated in the fusion, a total of 880 wells showing growth were screened, with a total of 94 X series and 24 I series clones selected for further work. Analysis of the fusion via Poisson distribution, suggested that there was a 4.6% probability that any well showing growth contained 2 or more clones, or 5 to 6 of the total clones being multiclonal. Of the 1 18 clones selected, 37-X and 8-1 series were eventually lost due to instability or lack of growth without feeder cells. A total of 32 subclones were selected based on selectivity of antibody binding to cancer positive urines versus the normal urines and on retention of assay OD with dilution of culture supernatant to select for high affinity and good production level. Samples of spent culture media from the following clones were evaluated for their potential utility in a clinical assay for the antigen described in Example III: 1-7.3, 1-8.2, 1-10.2, 1-11.1, 1-12.2, 1-17.3, X-4.1, X-13.1, X-13.2, X-22.2, X-28.1, X-44.1, X-48.1, X- 49.1, X-49.2, X-50.3, X-52.1, X-53.2, X-55.1, X-56.3, X-59.1, X-60.2, X-61.2, X-62.1, X-63.2, X-64.3, X-67.2, X-69.1, X-70.2, X-84.2, and X-87.2. A preferred monoclonal antibody pair for assays is X-13.2 (conjugate MAb) and X-52.1 (capture MAb). EXAMPLE II DEVELOPMENT OF GOAT POLYCLONAL ANTIBODIES A. Goat Immunization 1. Antigen Heparin-Agarose chromatography (Example I.A., above) fractions from three TCC-positive patients were pooled and dialysed against phosphate buffered saline (PBS). Protein concentration was determined to be 2 mg/mL. Thimerosal was added to a final concentration of 0.02%, and 0.25 mL aliquots were frozen until use. Table 1 is a listing of the amounts and references of the antigens comprising Pool I. Table 1 Antigen Pool I Patient ID Date of Sample Fractions Protein Cone. Volume Used 1 3/29/94 " 13-31* - 1 3/30/94 14-30* 3.2 mg/mL* 0.5 mL 2 6/6/94 11-19 0.91 mg/mL 1.5 mL 3 6/8/94 11-24 3.6 mg/mL 0.7 mL * both sets of fractions were combined before protein determination. 2. Immunization For immunization, a vial of antigen was thawed and mixed. An aliquot of 0.125 mL of antigen was mixed into 0.75 mL of PBS, and drawn into a 5 mL glass syringe. This syringe was attached to another such syringe containing 1 mL of Difco Freund's adjuvant, via a double-hub emulsifying needle. The first immunization was with Freund's Complete adjuvant, all others were with Freund's incomplete adjuvant. The emulsion was formed by alternately forcing the total mixture from one syringe to the other. The stability of the emulsion was tested by removing one syringe from the needle and dipping the end into a beaker of tap water. If a small amount of emulsion expressed into the water did not immediately spread over the surface, the emulsion was deemed stable. The entire amount of emulsion was collected into one syringe, which was capped and stored on ice until used. Total protein in the inoculum was 0.25 mg. Goats (R & R Rabbitry, Inc., Stanwood, WA) were 5 1/2 months of age and weighed approximately 34 kg. when the first immunization was administered. The second and third immunizations were given thirty and sixty days later. B. Antibody Analysis Serum samples were taken pre-immunization and two weeks after the second and third immunizations and were analyzed via ELISA using the antigen coated onto microplates. The assay was similar to the ELISA used for the mouse serum titer with the exception that antigen Pool I and rabbit anti-goat IgG-HRP were used and the dilution range employed was from 6 x IO 3 to 1.861 x IO 6 . The results of the assay were the following: 1. Pre-immunization sample showed no antibody titer as expected. 2. Samples after the second and third immunizations showed a maximal OD at about 3x10 3 dilution, a half maximal signal at about 8 x 10 4 and background at 1 x 10 6 . The signal at a dilution of 1 x 10 5 was 1.4. 3. In another experiment, cross reactions were tested and rated on a scale of 0 to 4 at a serum dilution of 1 x 10 5 , and were as follows in Table 2, with 4 being highest OD: Table 2 human Collagen Type IV 0 human Fibronectin 1 human Laminin 0 human Fibrinogen 1 Bovine submaxillary mucin 0 human red blood cells 0 Pool I 4 (rabbit anti-goat IgG-alkaline phosphatase was substituted for the rabbit anti-goat IgG-HRP). EXAMPLE III PURIFICATION AND CHARACTERIZATION OF ANTIGEN A. Purification 1. Heparin-Agarose Chromatography of Urine Twenty-four hour urine samples were centrifuged in a Beckman centrifuge, Model #J2-21, S/N 5539, using a JA-10 rotor at 6,000 φm for 20 minutes. The clarified urine sample was then concentrated using an Amicon stirred cell, 76 mm, (cat# 5124) fitted with a YM30 membrane MWCO 30,000 daltons (Amicon, cat# 13742) or a Microgon hollow fiber concentrator, 50,000 MWCO (cat# Ml 5S-260-01N) to approximately 100X concentration. The concentrated sample was diluted 1 :1 with 25 mM Tris-HCl, pH 7.4, and loaded onto a column of Heparin- Affigel (BioRad, cat# 153- 6173), equilibrated in 25 mM Tris-HCl pH 7.4, at a flow rate of 2.0 mL/min. The sample was followed with equilibration buffer until the A280 elution profile returned to background. Bound material was eluted with a 100 mL, linear NaCI gradient from 0 to 250 mM NaCI in 25 mM Tris-HCl, pH 7.4, followed by 50 mL of 250 mM NaCI, 25 mM Tris-HCl, pH 7.4, and finally 20 mL of a 1 OX PBS, pH 7.4, solution. Five mL fractions were collected and fractions from the trailing half of the elution peak were pooled. Pooled fractions were concentrated with an Amicon stirred cell, 43 mm (cat# 5122), fitted with a YM30 membrane, MWCO 30,000 daltons (cat# 13722). 2. Protein A Chromatography of 24 Hour Urine Protein A Chromatography was performed on a 24 hour urine from a TCC+ patient to determine whether this tumor antigen could be part of an immune complex. The urine (6 mL) was diluted to 12 mL with the addition of 6 mL of 20 mM sodium phosphate, pH 7.4. The diluted urine (7.3 mL) was loaded on a 1.0 mL Protein A cartridge (BioRad, Richmond, CA, cat# 732-0093) equilibrated in 20 mM sodium phosphate, pH 7.4, at 0.5 mL/min. The flow through volume plus 5.0 mL of wash buffer was collected and labeled as "flow through" (total volume = 12.3 mL). Bound material was eluted with 100 mM citrate buffer, pH 3.0, and neutralized immediately with the addition of 100 μl of a 1.0 M Tris-HCl, pH 8.0, to each 3 mL fraction. Eluted antigen was pooled (~ 6 mL) and the sample load, flow through, and eluted pool, at dilutions of 1 :20 to 1 :2560, were tested in the double monoclonal microtiter plate assay described in detail below (Example IV.B.). Approximately 97.5% of the activity loaded was contained within the flow through peak. The 2-3% activity in the eluted pool was probably due to incomplete washing. Thus, this antigen is not part of an immune complex involving IgG, and the use of immobilized Protein A would not be effective in extracting the antigen from specimens. 3. MAb 13.2 and MAb 52.1 Affinity Chromatography of 24 Hour Urine Aliquots of 24-four hour urines were diluted 1 : 1 with 25 mM Tris-HCl, 250 mM NaCI, pH 7.4, and loaded onto 5 mL MAb affinity columns (BioRad AlO gel) prepared with MAb X-13.2 or MAb X-52.1 (Example I.F.). To serve as a control for urine materials binding non-specifically to IgG, an AlO control column was prepared using Protein A-purified, normal mouse serum. Samples were loaded at 0.5 mL/min. The sample was eluted with 25 mM Tris-HCl, 250 mM NaCI, pH 7.4, until the OD280 baseline was reached. The bound material was then eluted with 100 mM glycine-HCl, pH 3.0. The eluted fractions, 5 mL each, were collected in tubes containing 100 μl of 1.0 M Tris-HCl, pH 8.0. Purity was assessed by SDS-PAGE using Novex 8-16% and or 4-12% polyacrylamide gels under reducing and non reducing conditions, along with Novex (San Diego, CA) Mark XII molecular weight standards (6 to 200 kD). The gels were stained with Coomassie Blue R250 followed by silver staining and scanned using a BioRad GS7000 densitometer. Molecular weights of individual bands are estimated based on the Rf values of the molecular weight standards (Example III.C). B. Native Molecular Weight Estimation of the Antigen bv Gel Filtration A gel filtration column was prepared with Pharmacia Sephacryl S-300 (Pharmacia, Piscataway, NJ, Cat# 17-0599-01). Briefly, deionized water is added to S- 300 gel to form a 50% slurry and added to a 1.0 L vacuum flask. The slurry was 32 degassed by gently swirling the slurry while pulling a vacuum. The degassed gel was then poured into a 2.6 x 100 cm, Pharmacia XK glass chromatography column (cat# 19- 0147-01) and packed at a 4.0 mL/min flow rate. The column bed was then equilibrated with 10 to 20 volumes of PBS containing 0.05% sodium azide and fitted with a Pharmacia column flow adapter (cat# 19-0688-01 ). A set of gel filtration standards (BioRad cat# 151-1901), with a range of 1.3 to 670 kD was dissolved in column equilibration buffer (PBS), filtered through a 0.45 μm Acrodisc, and loaded onto the column at 0.7 mL/min. The elution profile of the absorbance at a 280 nm wavelength was recorded at 2.0 cm/hr. A high molecular weight aggregate eluted at the column void volume (Vo). Each protein peak had its elution volume (Ve) determined by multiplying the time of elution of the maximum absorbance by the flow rate. A linear calibration curve was generated by graphing the Ve/Vo of the standard proteins vs. the logs of their molecular weights. Molecular weight estimates of the samples' peaks were made using the linear equation generated by the calibration curve. A twenty-four hour urine from a TCC+ patient was concentrated using an Amicon stirred cell, fitted with a 43 mm, YM30 membrane (MWCO 30,000 dalton) (cat# 5122). The urine was concentrated 3 OOX to ~ 0.5 mL and loaded onto the XK S300 column at 0.7 mL/min and 7 minute fractions were collected. The individual fractions were tested in the double monoclonal assay (described in Example IV.B.) to detect the presence of the antigen. A range of native molecular weights for the active fractions was calculated as 267 kD to 395 kD. C. Subunit Molecular Weights of the Components of Affinity-Purified Antigen bv SDS-PAGE MAb X-13.2.1 affinity purified urine antigen (Example III.A.3.) was applied, in the presence or absence of dithiothreitol (DTT; reducing sample buffer), onto a Novex discontinuous, 8% polyacrylamide, two well gel. A reference well contained Novex Mark XII SDS-PAGE molecular weight (MW) standards. Gels were electrophoresed at 125 V constant until the sample front reached within ~0.5 cm of the bottom of the gel. The gel's protein bands were then transferred to PVDF and stained with Coomassie Blue R250. Rf values were calculated for the Mark XII individual molecular weight standards by dividing the distance the band moved through the resolving gel by the distance of the sample front from the top of the resolving gel. A linear standard curve was established by plotting Rf values versus log MW for each MW standard. Sample bands' molecular weights were estimated by calculating their Rf values and entering these values (yi) in the standard curve equation. Immobilized MAb X-52.1 bound approximately 10 components with apparent molecular weights 151, 130, 87, 77, 60, 49, 39, 29, 25, and 10 kD under reducing conditions (i.e., in the presence of DTT). Only bands at 151, 130, and 39 kD appeared to be specific for the MAb X-52.1 in that the other proteins also bound to immobilized non-specific mouse IgG. Of these bands, that corresponding to a molecular weight of 151 kD is typically the most intensely staining. In contrast, immobilized MAb X-13.2 affinity purified fractions were generally cleaner than those obtained with immobilized MAb X-52.1, containing predominant bands at 151, 130, and 39 kD with only very minor contaminant bands at 77, 60, and 25 kD. Under non¬ reducing conditions, the monoclonal antibody-specific bands exhibited apparent molecular weights of approximately 138 kD, 121 kD, and 40 kD, with the 138 kD band being typically the most intense. The shift in apparent molecular weights of the dominant band from 138 kD to 151 kD upon reduction and the 121 kD band to 130 kD upon reduction could be due to the presence of a large number of intra-chain disulfide bonds in these molecules. This characteristic electrophoretic behavior formed the basis for the antigen assay described in Example IV.F. D. Western Blot Analysis of Partially Purified Antigen Preparations The urine samples purified on Heparin-Agarose (Example III .A.I .) were diluted 1:2 with SDS-PAGE 2X Sample Buffer (Novex, cat# LC 2676) in the presence dithiothreitol and heated at 100°C in a boiling water bath for 5 minutes, then allowed to cool to room temperature. The sample preparations were loaded onto an 8-16% acrylamide, 1.0 mm thick, 10-well, discontinuous Novex SDS-PAGE gel (Novex, San Diego, CA, cat# EC6045) and electrophoresed at 125 V constant for 190 V-h using a Novex electrophoresis chamber (Novex, cat# EI9001) and a BioRad Power Unit 500V (cat# 165-4710). Novex SeeBlue Molecular Weight Standards (cat* LC5625) were loaded into a reference well. The SDS-PAGE bands were transferred to PVDF paper (Novex, cat# LC2002) in Novex Transfer buffer (cat# LC 3675) using a Novex Transfer apparatus (Novex, cat# EI9051) and BioRad 500 power supply at 125 mA constant for 60 minutes. The PVDF paper was blocked with PBS containing 2% non-fat dry milk solution for 60 minutes, washed with PBS, containing Tween-20 (0.05%). MAbs are diluted in PBS-Tween to 2 μg/mL were added to the PVDF paper for 2 hours, washed with PBS-Tween, and incubated with an anti-mouse IgG alkaline phosphatase conjugate for 1 hour. The PVDF was washed with 50 mM Tris containing 5 mM MgCl 2 and then a NBT/BCIP substrate solution was added in the 50 mM Tris-5 mM MgCl 2 solution and the bands were developed. MAb 13.2 showed three major bands at 138, 121 , and 40 kD. MAb 52.1 recognized the same bands, but reacted with the 40 kD band to a greater extent and with the 121 kD band to a lesser degree than MAb X-13.2. E. Amino Acid Sequence Determinations 1. N- Terminal Amino Acid Sequence A 450 mL pool of three urines was initially chromatographed on a Heparin- Affigel column (2.5 x 16 cm), as described in Example III.A.l. The antigen containing fractions were eluted with 100 mM NaCI in 25 mM Tris-HCl buffer, pH 7.4, were pooled and 50 mL at 265 μg/mL was loaded directly onto a monoclonal affinity column, 2 mL, prepared with MAb X- 13.2.1 (Section II.F.) and Affigel 10 (BioRad, cat# 153-6099). The bound antigen was eluted with a 100 mM sodium citrate, pH 2.5, and immediately neutralized with a 1.0 M Tris-HCl buffer, pH 8.0. The MAb affinity purified antigen was diluted 1 : 1 with SDS-PAGE 2X Sample Buffer (Novex, cat# LC 2676) containing 2% DTT and heated at 100°C in a boiling water bath for 5 minutes, then allowed to cool at room temperature. The sample preparation was loaded onto an 8% acrylamide, 1.0 mm thick, 2-well, discontinuous Novex SDS-PAGE gel (Novex, cat# EC6012) and electrophoresed at 125 V constant for 190 V-h using a Novex electrophoresis chamber (Novex, cat# EI9001) and a BioRad Power Unit 500V (cat# 165-4710). BioRad SDS-PAGE Molecular Weight Standards (cat# 161 -0317) were loaded into a reference well. The gel was removed and placed in a container of 10 mM CAPSO buffer, pH 9.0, containing 0.05% SDS on a rocker platform while the gel transfer sandwich was prepared. The SDS-PAGE bands were transferred to PVDF membrane (Novex, cat# LC2002) using a Novex Transfer apparatus (Novex, cat# EI9051) and BioRad 500 power supply at 125 mA constant for 60 minutes. The PVDF membrane was removed and rinsed with deionized water and stained in a 0.1% Coomassie Blue R- 250 in 20%) methanol protein staining solution for approximately 10 minutes. The stained PVDF was then destained with several changes of 30% methanol until the background stain was minimal, and was followed by extensive washing in deionized water. The PVDF membrane was then allowed to dry at room temperature on a paper towel. The stained bands of interest were excised with a clean razor blade and placed in capped tubes. The samples were carried to the University of Washington (Seattle, WA) for sequencing by Edman degradation. The principal amino acid sequence thus obtained was: E D C N ? L P P R ? N T (SEQ ID NO:l), where the symbol "?" indicates a residue which could not be identified. 2. Trypsin Digestion: Internal Amino Acid Sequence A small amount (50-100 μL) of immobilized trypsin (Pierce Chemical Co., Rockford, IL) was added to a 600 μL Eppendorf tube along with an equal volume of PBS. After gentle mixing, the slurry was spun down at 10K φm for about 30 seconds. The supernatant solution was pipetted off and two times the slurry volume of PBS was added, mixed and the spin repeated. This wash step was repeated twice more and the slurry brought back to the original volume with PBS. A known quantity of antigen was added to a clean 600 μL Eppendorf tube and PBS added to bring the concentration to 0.5 mg/mL. Immobilized trypsin was added in a 1 :10 E/S ratio (w/w), and the solution was gently mixed and placed on a rotator for four hours at room temperature (21 -23°C). Digestion patterns were visualized by SDS-PAGE using a 4-12%) gradient Tris-Glycine precast gel with Tris-Glycine SDS running buffer (NOVEX, San Diego, CA). Samples were mixed with the appropriate amount of 4x sample load buffer, containing 4% dithiothreitol (ACS grade reagent), boiled for two minutes and then loaded on the gel. Gels were run for 240 Vhr at 125 volts constant, stained in 0.01% Coomassie R250 (BioRad, Hercules, CA,) in 10% acetic acid, 50% methanol for an hour and destained in 10% acetic acid, 50% methanol for about 20 minutes. Gels were transferred into water for one hour, into GelDry solution (NOVEX, San Diego, CA) for twenty minutes and then dried in between two sheets of DryEase mini precut cellophane (NOVEX, San Diego, CA) overnight. Seven fragments were observed. Material for sequencing was prepared by digestion with trypsin as described above. Approximately 300 μg of urine antigen was digested, boiled in 1/3 volume load buffer with 4% DTT and electrophoresed on a 10% Tricine gels (NOVEX, San Diego, CA). The gels were rinsed (about 1-2 minutes each time) in three changes of 10 mM CAPS (Sigma Chemical Co., St. Louis, MO) containing 10% methanol (ACS grade reagent), pH 11.0 (blotting buffer) to remove any contaminants from the gels. The proteins were transferred onto PVDF membranes (NOVEX, San Diego, CA) by electroblotting in blotting buffer at 30 V constant for 1.0 hour. After blotting, the membranes were washed once with fresh blotting buffer and then stained for two minutes with 0.1 % Coomassie R250 in 10% acetic acid containing 50% methanol. The blots were destained in 10% acetic acid with 50% methanol for approximately 15 minutes, rinsed three times with deionized water and then air dried. Stained portions of the blots were excised, placed in 15 mL conical tubes and stored at -20°C until sequenced. Sequencing was performed at the laboratory of Dr. Ken Walsh, Department of Biochemistry, University of Washington (Seattle, WA), and the results are shown in Table 3. Residues tentatively assigned by the operator appear in parenthesis and unassignable residues are indicated by a question mark. Table 3 Sequences of Urine Antigen Tryptic Fragments Sample 1 Fragment (~MW) Sequence SEQ ID 1 128 kD GPYFPVAVGKYY?(Y)Y?D NO 2 sequence starts at CFH AA 324 [RPYFPVAVGKYYS Y YCD] NO.12 2 79 kD RPYFPVAVGKYYS?Y?DE?F???S NO:3 sequence starts at CFH AA 324 [RPYFPVAVGKYYSYYCDEHFETPS] NO:13 3 46 kD SSQESYAHGTK N0:4 sequence starts at CFH AA 868 [SSQESYAHGTK] N0:4 4 37 kD EDCNELPP?RNTEIL?GSW-D NO.5 sequence starts at CFH AA 1 [EDCNELPPRRNTEILTGSWSD] NO: 14 5 66 kD RPYFPVVAVGKYYSYY?DEHFE?P N0:6 sequence starts at CFH AA 324 [RPYFP-VAVGKYYSYYCDEHFETP] NO:15 6 33 kD 2 SLGNVIMV7RKGEWVALNPLRK N0:7 sequence starts at CFH AA 40 [SLGNVIMVCRKGEWVALNPLRK] NO:16 sequence starts at CFH AA 324 3 RPYFPVAVGKY N0:8 [RPYFPVAVGKY] N0:8 * Arm no aci d residue numbers refer to the mature CFH molecule 2 Major protein sequence 3 Minor protein sequence The similarity of the partial amino acid sequences of the antigen with those of the reported sequence for human complement Factor H (shown in brackets) demonstrates that the antigen detected is a member of a complement Factor H-related family of proteins as disclosed herein. F. C3b - Decay Accelerator Activity of the Antigen C3b was prepared from C3 (Sigma Chemical Co., St. Louis, MO) by trypsin digestion, using immobilized trypsin (Pierce Chemical Co., Rockford IL), and an enzyme to substrate (E/S) ratio of 1 :25 at room temperature for fifteen minutes. The digest was spun at 1 Ok φm for about 30 seconds to pellet the enzyme, and the supernate removed. The supernate was checked for the presence of C3b by SDS-PAGE under reducing conditions. Five μg each of affinity-purified antigen from either urine or serum, 5 μg Factor I (Sigma Chemical Co., St. Louis, MO) and 50 μg C3b (Sigma Chemical Co., St. Louis, MO) were incubated with gentle mixing at 37°C for 90 minutes. Six μl of undiluted, dialyzed patient urine samples, 3 μl Factor I (FI) and 30 μl C3b were incubated as described above in a separate experiment. A small portion of each reaction mixture was boiled for two minutes with 1/3 volume 4x load buffer, 4% DTT and loaded onto a 4-12% Tris-Glycine gradient gel. Gels were run for 240 Vhr at 125 volts constant, stained in 0.01% Coomassie R250 in 10% acetic acid, 50% methanol for an hour and destained in 10% acetic acid, 50% methanol for about 20 minutes. Gels were transferred into water for one hour, into GelDry solution (NOVEX, San Diego, C A) for twenty minutes and dried between two sheets of DryEase mini precut cellophane (NOVEX, San Diego, CA) overnight. The dried gel was analyzed with a BioRad Model GS-700 Imaging Densitometer equipped with BioRad' s Molecular Analyst software and the molecular weights of the digestion fragments were estimated using Mark 12 molecular weight markers as standards. The disappearance of the C3b α' fragment at a molecular weight of 108,000 daltons and the concurrent appearance of fragments with molecular weights of 67,000, and 47,000 daltons indicate the digestion of C3b by Factor I. This digestion only occurs when mediated by a cofactor molecule, such as Complement Factor H, as was apparent in the control runs. For example C3b, when incubated with Factor I alone, does not degrade. Urine affinity purified antigen mediated the digestion of C3b by Factor I, indicating that this antigen has a functional C3b binding site and acts as a cofactor in the digestion of C3b by Factor I. TCC + urine samples also functioned as cofactor for the digestion of C3b by Factor I, while a normal urine sample did not. EXAMPLE IV ASSAYS FOR THE ANTIGEN Given the characteristics of the antigen as described above and given the disclosure herein for generating and selecting antibodies and the development of certain assays described herein to detect the antigen, a number of additional assay formats beyond those described herein for this antigen may be readily developed by those of ordinary skill in the art. Suitable assay formats include competitive formats, sandwich formats (Examples IV.A., IV.B. and IV.C), assays based on the biological or chemical properties of the antigen (Example IV.D. and IV.E.), assays based on the simultaneous binding of the antigen to a specific macromolecule (e.g., C3b) and to a monoclonal antibody (Example IV.D.), assays based on the appearance of a band of appropriate size in partially-purified specimens (Example IV.F.), and RT-PCR (Example IV.G.). A preferred format involves sandwich immunoassays and the most preferred employs a monoclonal antibody immobilized on a solid surface and a second monoclonal antibody, which recognizes an epitope distinct from that of the first, conjugated to a detection agent. That detection agent could be an enzyme (Example IV.B.), colloidal gold (Example IV.C), or any of a number of other such agents known to those of ordinary skill in the art. These include fluorescent molecules, radioisotopes, and biotin (which would subsequently bind to avidin or strepavidin-labeled detecting agent). A. Identifying Potential Antibody Pairs Definitions for the section: Indirect Assay Format: Antigen coated on plate; reaction with MAb; signal generation by Goat Anti-mouse conjugated to alkaline phosphatase (GAM-AP). Direct Assay Format: Antigen coated on plate; reaction with and signal generation by specific MAb-AP. Sandwich Assay Format: As usual Initial screening of the cell culture supematants (Example I.F.) was carried out using an ELISA in an indirect format. The assay consisted of the following in order: (1) diluted urine samples were adsorbed on a microtiter plate; (2) following washing, the microtiter plate wells were incubated with supematants of cell cultures of the clones of interest; (3) following another wash, the plates were incubated with alkaline phosphatase-conjugated goat anti-mouse IgG; (4) following a final wash, the plates were incubated with p-nitrophenyl phosphate substrate (pNPP); and, finally, (5) the reactions were stopped by addition of concentrated EDTA to each well and the color measured at a wavelength of 410 nm on a microplate reader. Cell culture supematants from 32 different clones (Example I.F.) were tested against single dilutions of urine samples in the first stage of screening of this nature. The urine samples used during this first stage consisted of eight from normal individuals and eight from individuals with transitional cell carcinoma of the bladder (TCC) of various stages and grades. Still using the indirect format, supematants from 23 clones which showed acceptably low reactivity with the normal samples (specificity = 7/8 or 8/8) and generally positive reactivity with the TCC urine samples (between 6/8 and 8/8) were then further tested against serial dilutions of eight urine samples from patients with TCC of various stages and grades. Based on their behavior in this experiment with serially diluted urine samples adsorbed to the plates, twelve were selected for further study. In this experiment, plates were prepared with fixed dilutions of eight urine samples and serial dilutions (between 1/10 and 1/1280) of each of the antibodies were applied. All twelve of these showed sufficiently high reactivity that the eight TCC-positive samples were serially diluted and again assayed. The final experiment using the assay in the indirect format consisted of testing the twelve cell culture supematants against twelve TCC-positive urine pools. Based on these initial experiments in the indirect format, all twelve antibodies were selected for further testing/screening utilizing the sandwich format of the assay. Initial testing of antibodies conjugated to alkaline phosphatase (AP), as described in Example IV.B.2., was carried out utilizing an assay in the direct format as follows: (1) diluted urine samples were adsorbed on a microtiter plate; (2) following washing, the plates were incubated with AP-conjugated antibodies from specific clones; (3) following a final wash, the plates were incubated with pNPP; and, finally, (4) the reactions were stopped and measured as above. Based on the results obtained from seven conjugates tested on a small number of urine samples in this manner, all seven were selected for further study in the sandwich format of the ELISA. Thirteen monoclonal antibodies (Example I.F.) and one goat polyclonal preparation (Example II) were tested as capture antibodies in combination with the seven alkaline phosphatase conjugates in the sandwich ELISA format as follows: (1) individual capture antibodies were adsorbed on microtiter plates; (2) following washing, diluted urine samples were added to the wells and incubated to allow binding of the antigen to the antibody; (3) following another wash, single conjugates (as described in B. above) were added to individual wells and incubated to allow binding to the antibody-bound antigen, if present; (4) following a final wash, the plates were incubated with pNPP; and, finally, (5) the reactions were stopped and measured as above. A total of 107 potential antibody pairs were first tested against one normal and seven TCC-positive urine samples. From these, a selection of 33 pairs were chosen to be tested against an expanded series of urines from 31 patients and one normal individual. From the results of this testing, seven antibody pairs were selected for further testing against a much expanded selection of 120 patient urine samples, but including also 20 samples from normal individuals. From this extensive testing of these seven pairs, a single monoclonal antibody pair (X52.1/X13.2-AP) was selected as the most preferred on the basis of (1) its positive response with the greatest number of samples from TCC-positive patients, (2) its negative response with the greatest number of samples from non-TCC-positive patients, and (3) low nonspecific reaction with urine samples from normal, non-diseased individuals. In addition, an alternative antibody pair was selected (X52.1/X62.1-AP). B. Sandwich ELISA The sandwich ELISA, utilizing the most preferred pair as selected above, was further optimized with respect to the following items: (1) coating level of capture antibody; (2) concentration of conjugate; (3) enzyme-to-antibody ratio in the conjugate; (4) reaction kinetics/incubation times; (4) composition of assay and wash buffers and of conjugate and specimen diluents; and (5) formulation of standards and controls. The assay as optimized is performed as follows: 1. Preparation of Coated Plates The plates were coated with 150 μl per well of monoclonal antibody at a concentration of 5μg/mL in carbonate buffer at pH 9.6. The plates were then blocked with 2% bovine serum albumin in phosphate-buffered saline at pH 7.4, followed by blocking with 4% sucrose. The sucrose solution was decanted, and the plates were dried overnight at room temperature. 2. Preparation of MAb-Alkaline Phosphatase Conjugates Antibodies were purified by chromatography on immobilized Protein G or Protein A by standard techniques. Although antibody-enzyme conjugates could be prepared using a variety of coupling techniques (for review see Scouten, W.H., Methods in Enzymology 135:30-65, 1987), a minor variation of a method described by S. Hashida and E. Ishikawa (Anal. Lett. 18, 59:1 143-1155, 1985) was used. Briefly, purified monoclonal antibodies were treated with excess N-acetylhomocysteine thiolactone (AHTL) at neutral pH to introduce reactive thiol groups, and then desalted to remove excess AHTL. Separately, alkaline phosphatase (AP) was treated with excess sulfosuccinimidyl 4-(N-maleimido-methyl) cyclohexane- 1 -carboxylate to introduce maleimido groups, and excess reagent was removed by desalting. The conjugates were prepared by mixing antibody and enzyme derivatives, which became covalently coupled via thioether bonds. Any excess maleimido groups were then capped by reaction with cysteamine. 3. Assav Format A volume of 175 μl of assay buffer was pipetted into each well to be utilized in carrying out the assay. The buffer was followed by 25 μl of samples, standards, or controls, thus yielding a 1/8 dilution in the well. Incubation of the covered plate was performed at 37°C for 60 minutes. Following washing, 200 μl of working dilution of conjugate was added to the aspirated well. The covered plate was again incubated for 60 minutes at 37°C. Following a final wash, 200 μl of pNPP substrate was pipetted into each well, and the covered plate was incubated at 37°C for 30 minutes. After pipetting, 50 μl of stop solution into each well, the reaction mixtures in each well were measured at 410 nm. 4. Typical Results Eighty seven urine samples were assayed by the ELISA using the format described above. These samples included 23 clinical specimens taken from patients diagnosed as currently having transitional cell carcinoma (TCC) and 64 others. The results are tabulated below in Table 4. Sensitivity is reported as the percentage of specimens from TCC-positive patients that correctly produce a positive result in the assay. Specificity is reported as the percentage of urines from individuals without TCC that correctly produce a negative result in the assay. Table 4 Number Percentage of Specimens of Total Sensitivity 23 48% Specificity Healthy 25 88% Non-GU Disease 15 87% GU Malignancy 3 100% Other GU Disease 10 70% Chronic Inflammation 11 27% (Urinary Tract) A graphical representation of the data for each specimen, expressed as Units of analyte/mL, is given below. (Note that the categories correspond to those specified in the above table and that GU=Genitourinary.) NON-GU DISEASE GU MALIGNANCY & OTHER GU DISEASES Sample Number Sample Number CHRONIC INFLAMMATION Sample Number It is clear that the ELISA described here for the detection of this tumor antigen yields a positive reaction with a significant number of urine specimens taken from patients diagnosed with bladder TCC. Samples which yield negative results, although taken from TCC-positive patients, correspond to those with disease at an early stage and/or of low grade. With respect to those patients identified as having chronic inflammation, several have a history of TCC. C. Rapid Assay Monoclonal antibodies specific for the antigen (Example I.F.) were utilized in a lateral flow format to produce a qualitative assay for bladder cancer using urine as the specimen. The lateral flow format consisted of a colloidal gold antibody conjugate and an immobilized capture antibody on a nitrocellulose membrane. Upon interaction of the urine sample with the colloidal gold conjugate, the antigen in the urine sample formed an antigen-antibody conjugate complex. This complex migrated by capillary flow through the membrane and contacted the immobilized anti-antigen capture antibody (test zone). The capture antibody bound the antigen-antibody conjugate complex, forming a visually detectable colored signal in the test zone. Material not bound by the capture antibody continued to migrate through the membrane and contact an immobilized goat anti-mouse antibody (control zone) which bound the colloidal gold conjugate regardless of the presence of antigen, forming a visually detectable signal in the control zone. Purified monoclonal antibody X-13.2 was conjugated to colloidal gold according to Frens (Frens, G., Nature, Phys. Sci. 241:20-22, 1973). Briefly, the gold sol was prepared by reduction of tetrachloroauric acid by trisodium citrate. The solution was boiled until a color change was observed. MAb XI 3.2 was adsorbed to the gold sol at 0.3 mg/ml for 5 minutes at pH 9. The conjugate was blocked with 0.5% BSA and washed twice with the conjugation buffer. The washed conjugate was then diluted 7- fold into 2% BSA with 50mM Tris, pH 9, and 0.05% NaN 3 . The washed conjugate was used to saturate strips (10.5 x 0.25 in.) of glass fiber mesh (Lydall, Hamptonville, NC). These conjugate strips were then dried overnight under reduced pressure at ambient temperature. An airbrush sprayer was used to immobilize the capture and control antibodies on the membrane. Purified monoclonal antibody X-52.1 at 2 mg/ml was sprayed as a line onto a section of nitrocellulose membrane (8 μm pore size, 50m x 1 inch, Whatman, Fairfield, NJ) approximately 3/8 in. from one edge of the membrane strip. A goat anti-mouse antibody (Chemicon, Temecula, CA) at 2 mg/ml was sprayed approximately 3/8 in. from the other edge of the membrane. The membrane was then dried overnight under reduced pressure at ambient temperature and cut into 10.5 in. strips. The 10.5 in. strips of coated glass fiber mesh and sprayed membrane were then assembled onto plastic cards (polypropylene, 10.5 x 2.25 in.) using double- sided tape (below). The membrane was first placed near the center of the plastic card. The conjugate pad was then placed so as to overlap the membrane on the proximal side (near the x52.1 immobilized antibody) of the membrane and an absorbent cotton paper strip (Whatman, 10.5 x 0.75 in.) was placed so as to overlap on the distal side (near the goat anti-mouse immobilized antibody) of the membrane. Finally, a second absorbent cotton paper strip was placed in overlapping contact with the conjugate pad to accept the sample. The assembled cards were cut crosswise and the resulting small strips placed into plastic housings which provided a well for containment of the sample and a viewing window in the nitrocellulose region for reading the results. Configuration of lateral flow assav components: Positive Negative 1. Results Rapid assays were carried out by placing 250 μl of patient urine in the sample well. After 10 minutes, the results were read. A positive result will show a pink-puφle line in the test zone (zone of immobilized X-52.1) and a pink-purple line in the control zone (zone of immobilized goat anti-mouse). A negative result will show no line in the test zone and a pink-puφle line in the control zone. The absence of a line in the control zone indicates that the reagents in the test did not function properly and this test is invalid. Twenty three TCC-positive clinical specimens and 64 other urine specimens were assayed in the lateral flow assay. The results are given in Table 5 below. Sensitivity is reported as the percentage of TCC-positive specimens that correctly produced a positive result in the lateral flow assay. Specificity is reported as the percentage of TCC-negative urines that correctly produced a negative result in the lateral flow assay. Table 5 Sensitivity (23) 65% Specificity Healthy (25) 92% Non-GU Disease (30) 57% GU Malignancy (3) 100% Other GU Disease (6) 83% GU = Genitourinary From Table 5 it is clear that the bladder cancer lateral flow assay detects a large percentage of the TCC-positive specimens tested and distinguishes bladder cancer (TCC) from other normal and disease states. D. C3b-MAb ELISA 1. Method Immulon 4 (Dynatech, Chantilly, VA) microtiter strip wells were coated with 50 μl per well of 5 μg/ml trypsin treated C3 (converts C3 to C3b; see Example III.F. above for C3 source and method of activation) in 50 mM carbonate buffer, pH 9.6, either ovemight at 4°C or for two hours at 37°C. A control plate was coated with 50 μl per well of 2% BSA in PBS for two hours at 37°C. After a single wash with Tris-buffered saline (TBS) containing 0.1% Tween -20 (wash buffer), the plates were blocked with 100 μl per well of a 2% BSA solution in PBS for two hours at 37°C and washed four times. Antigen, diluted in assay diluent (1% BSA in TBS with 0.15M MgCl 2 , 0.15M ZnCl 2 ), was added at 50 μl per well and incubated for one hour at 37°C. The plates were washed four times and then the detection antibody (xl 3.2.1.1- alkaline phosphatase,) was applied at 0.25 μg/ml, 50 μl per well, and incubated at 37°C for 30 minutes. After four washes, 50 μl per well of p-nitrophenyl phosphate (Sigma, St. Louis, MO) at 1 mg/ml in IM diethanolamine (DEA) was added and the plate incubated for 30 minutes at 37°C. The reaction was stopped with 25 μl per well of stop solution (0.1 M EDTA, pH 9.8) and the plate read at 405 nm on a Dynatech MR7000 reader. 2. Results This assay format discriminated between TCC + urines (PES and CHD) and normal urines, yielding a positive signal for the TCC + urines. Dilutions E. C3b - Decay Accelerator Activity Assav The assay described above in the C3b decay accelerator activity section (Example III.F.) discriminated three patient urines (all TCC + ) from a normal urine pool. Therefore, it may be used as an assay to indicate bladder cancer in patients. F. SDS-PAGE Assav The antigen partially purified by heparin agarose chromatography (Example III.A.) may also be detected by SDS-PAGE under reducing and non-reducing conditions, since it displayed a characteristic apparent molecular weight shift upon reduction (Example III.D.). As described above, the reduced antigen exhibited an apparent molecular weight (as estimated by SDS-PAGE) of -151 kD as compared to -138 kD under non-reducing conditions, presumably due to numerous disulfide bridges. The antigen containing peak from heparin chromatography was diluted in 2X SDS- PAGE sample buffer with and without dithiothreitol (DTT) at 5%. The samples were heated for 5 minutes in a boiling water bath and then allowed to cool at room temperature. Aliquots (2 μL) were loaded onto an 8 well, 7.5% acrylamide Pharmacia PhastGel (cat# 17-0622-01) and run on the Pharmacia PhastSystem (cat# 18-1018-23) according to manufacturer's protocol. Reduced and non-reduced samples were run on the same gel and were separated by molecular weight standards and an empty lane loaded with non-reducing IX sample buffer. The gels were stained with Coomassie Blue R250 0.1% in 40% methanol and 10% acetic acid and then destained with 40% methanol with 10% acetic acid. Characteristic bands were seen on specimens with elevated antigen levels as detected by the ELISA (Example IV.B.). G. RT-PCR Assav 1. Cell Lines Several cell lines, particularly cell lines HTB-5 and HTB-9, which are derived from Transitional Cell Carcinoma (TCC) of the bladder and HeLaS3, which is derived from adenocarcinoma of the cervix (all from American Type Culture Collection, Rockville, MD), were tested to determine whether they produce mRNA coding for the antigen. Although the method selected for cell line analysis was RT- PCR (Reverse Transcriptase based Polymerase Chain Reaction amplification of messenger RNA, mRNA), a variety of procedures used to detect the presence of specific RNA can be used. Controls were performed using PCR target materials (the PAW 109 sequence) provided with commercial PCR kits, and its primers DM152 and DM151. Hybridoma cell line X-44.1 or normal human epithelial keratinocytes (Clonetics Coφ., San Diego, CA) were chosen as the irrelevant target (Negative controls). 2. Preparation of mRNA Preparation of mRNA was facilitated by the use of a Lysis Buffer containing: 7.5 M Guanidine HCI, 25 mM TES, 10 mM EDTA, 0.05% Taurodeoxycholate, 1 mM 2-mercaptoethanol, pH 7.5 (all reagents Molecular Biology grade from Sigma, St. Louis, MO.). This buffer eliminated the necessity for grinding or icing samples and resulted in a stable preparation of DNA and RNA. Cells were lysed in 1 mL lysis buffer per IO* cells/mL cell culture media (IMDM, Irvine Scientific; Irvine, CA) supplemented with 15% FBS (Hyclone; Logan, Utah). The lysate was extracted with equal volumes of phenol and chloroform/isoamyl alcohol. The aqueous phase was aspirated and re-extracted with an equal volume of chloroform /isoamyl alcohol. The aqueous phase was precipitated with 7/13 volumes 10M LiCl (all reagents Molecular Biology Grade from Sigma Chemicals, St. Louis, MO). The mRNA was prepared from the total RNA produced in the previous steps using a PolyATtract kit (Promega, Madison, WI). 3. RT-PCR Amplification RT-PCR amplification of antigen sequences was performed on a Perkin- Elmer 2400 Thermal Cycler using a GeneAmp PCR kit (Perkin-Elmer/Roche Molecular Systems, Branchburg, NJ). Amplification was performed with 3 μL purified mRNA, 35 cycles, for the first amplification step of each reaction. The RT primer was designated 753RT, sequence TCGTTCATTCTCCTTAT (SEQ ID NO:9). The PCR primer for the first reaction was designated 42M, sequence GCTGGTAAATGTCCTCT (SEQ ID NO: 10). For the nested PCR, 20 μL of product from the first PCR reaction was re- amplified for 35 cycles using the 753RT primer and primer 412M, sequence ATGTAATGAGGGGTATC (SEQ ID NO: 11). All primer concentrations were set at 0.2 μM, and the annealing temperature was set at 48°C. 4. Results Gel electrophoresis of the first-step RT-PCR products is shown in Figure 1. Analysis of the second-step products by electrophoresis in TAE buffer (40 mM Tris- acetate, 20 mM EDTA, pH 8.3) on a 1 percent agarose gel revealed a band of the expected size, 341 bp, upon staining with ethidium bromide (Figure 2, lane 3). Amplification of the kit positive control PAW 109 gave the expected 311 base pair product. Re-amplification of this product with the kit DM152 and 151 primers was negative, a common result when re-amplifying PCR product without changing primers. EXAMPLE V CERVICAL CANCER A. Cervical Specimen Handling and Disposal 1. Preparation of tubes for cervical sample collection and transportation: Tubes: Coming polypropylene, 4mls, cat # SP:T4188-5, containing 100 μl of saline solution each, and were shipped to the clinical sites. Cervical samples were collected using swabs. The swabs were inserted into the tubes and the tips broken off prior to covering the tubes for shipment. 2. Specimen Preparation prior to Assav: Add 2 mis of 100 mM NaCI, 10 mM Tris-HCl (pH 8.0) containing 1 mM EDTA to 2 tubes and this buffer containing 0.1% Tween to the third tube. (The third sample is for investigating sample elution from the swab and may be used for other extraction buffers as needed.) Vortex tubes for 3-5 minutes. Pull out the swab with forceps while squeezing to the side of the tube to let all fluids out of the swab. Recap the tubes and centrifuge for 5 minutes at 3000 φm in a table top centrifuge. Serially dilute the supernatant to 1/8 and 1/16 with buffer. Add 20 μl of protease inhibitor cocktail to each tube and 20 μl of the 10% sodium azide solution. Mix well and freeze remaining supematants at -80°C prior to testing in ELISA. B. ELISA Sixty-three cervical samples were tested in several EIA combinations (Example IV.B.) including the X-52.1MAb/X-13.1 ALP sandwich EIA. The following are the assay procedure and results: 1. Assav Procedure a. EIA plates (COSTAR-Hi binding) were coated with X-52.1 MAb at 5 μg/ml in Carb/Bicarb buffer (pH 9.6), 100 μl per well, and incubated ovemight at 4°C. b. Coating buffer was discarded and the plates were blocked with 250 μl per well with 2% BSA/PBS (pH 7.4) for 2 hrs at room temp, c. Plates were then washed once with TBS containing 0.1% Tween- 20. d. Cervical samples were thawed at room temperature, centrifuged at 3000 φm for 5 mins, then the supernatant was diluted to 1 :20 in 25mM Tris-HCl, pH 7.8, in tubes and 100 μl of sample/diluent transferred per well to duplicate wells. The plates were sealed with plate covers and incubated at 37°C for 2 hrs. e. Plates were washed 4x with TBS containing 0.1% Tween-20. f. X13.1-AP conjugate was diluted to 2 μg/ml in 1% BSA TBS (pH 7.4) and 100 μl transferred to each well. Plates were then covered and incubated at 37°C for 2 hrs. g. Plates were washed 4x with 0.1 %T/TBS. h. A 1 mg/ml solution of p-NPP/ IM diethanolamine was made (pH 9.8), and 100 μl was transfeπed to each well. Plates were sealed with plate covers and incubated at 37°C for 1/2 hour. i. Color development was stopped by adding 50 μl per well of 0.1M EDTA (pH 9.8). j . Plates were read at 410 nm on a Dynatech MR7000 2. Results Results from the study of the cervical specimens with this assay are tabulated in Table 6 and presented graphically below. Table 6 XB.lMAb/ X52.1- AP Sensitivity (cancers) n=15 73% Specificity (normals) n=19 100% % Adenocarcinoma above cutoff n=3 33% % Dysplasia above cutoff n= 19 16% % Atypicals above cutoff n=7 0% Cutoff (Mean OD of Normals +2SD) 0.201 CTA IN X52.1/X13.1AP AD iENO EXAMPLE VI PRODUCTION OF CFHRP IN CANCER A. Production of CFHφ in Cancer Cell Lines 1. Detection of Antigen in Cell Culture Media bv Immunossay Cell culture media were tested for the presence of antigen (complement Factor H-related protein, CFHφ) using the sandwich enzyme immunoassay as described in Example IV.B. The media tested were those taken from cell cultures used for the preparation of total cellular RNA. After removal of the cultured cells, the remaining media free of cells were then diluted, as necessary, and tested in the EIA, as described. Control experiments involved the testing of fresh media, in particular those specified by ATCC or Clonetics Coφoration (San Diego, CA) for the cell lines or primary cultures of interest. These were typically Modified Eagle's Media containing 10% fetal bovine serum (Sigma Chemical). 2. Detection of Message for Antigen in Cancer Cells bv RT-PCR cDNA was synthesized from mRNA present in preparations of total cellular RNA from cancer cell lines, using Reverse Transcriptase plus Random Hexamer primers. The concentrations of components within the reaction mixture were as follows: Component Final Concentration MgCl 2 5 mM KCl 50 mM Tris-HCl, pH 9.0 10 mM Triton X- 100 0.1% v/v dGTP I mM dATP 1 mM dCTP 1 mM dTTP I mM RNAsin l U/μL Random Hexamers 5 μM MuLV Reverse Transcriptase 2.5 U/μL Sample and DEPC-treated deionized water were added to bring the reaction volume to 20 μL. Between 1 and 5 μg of total RNA was added to each reaction mixture. Typically, 2 μg is sufficient. The cDNA reaction was allowed to proceed for 90 minutes at 42°C. PCR of huCFH and huCFHφ mRNA was performed primarily with primer pair 42M and 1040RT (TCTGGATAATCACAAGGTTTC) (SEQ ID NO: 17), and primer pair 2910M (GTCAGACAGTTATCAGTATGGAGAAGAAG) (SEQ ID NO:18) and 3610RT (CTGTTTGGCTGTCCACCTTAATGCTATG) (SEQ ID NO:19). In addition, the presence of the correct internal sequences were confirmed with the primer pair 410M (ACATGTAATGAGGGGTATCAA) (SEQ ID NO:20) and 1040RT. The PCR master mix consisted of the following: Component Final Concentration MgCl 2 3 mM KCl 50 mM Tris-HCl, pH 9.0 10 mM Triton X- 100 0.1% v/v Taq DNA Polymerase 2.5 U/100 μL Primer Pairs: 42M 1040RT 1-5 μM or 2910M/3610RT 1-5 μM or 410M/1040RT 1-5 μM PCR was performed by adding 80 μL of master mix to each cDNA reaction tube. Thermal cycling was performed in a Perkin-Elmer (Foster City, CA) model 2400 cycler for 40 cycles. Positive results were determined by electrophoresis (e.g., at 90 volts for 90 min.) on 2% agarose gels, followed by staining with ethidium bromide, and destaining in deionized water. For purposes of this application, moderate stringency hybridization and PCR amplification conditions are defined as those performed at the calculated melting temperature (Tm) of the probe/primer with the target. The recommended formula for calculating Tm, and its limitations, are well known in the art (i.e., are found in Sambrook, J., Fritsch, E.F. and T. Maniatis, Molecular Cloning, 2d Edition, Cold Spring Harbor Laboratory press, pp. 9.51-9.52, 1989). High stringency conditions are defined within this application as hybridization amplification performed at least 4°C above the calculated Tm. Using primer pair 42M/1040RT, CFHφ are detected at moderate (52°C) to high stringency (56°C) conditions, based on the homology of the cDNA to that of the primers identified in this application. Using primer pair 2910M/3610RT, CHFφ are detected at moderate (67°C) to high stringency (72°C) conditions, based on the homology of the cDNA to the primer pair. For internal cDNA sequence, using the primer pair 410M/1040RT, CFHφ are detected at moderate (48°C) to high stringency (56°C) conditions. For PCR studies, DEPC-treated deionized water, Taq polymerase. RNAsin, and MuLV Reverse Transcriptase were from Promega Coφoration (Madison. WI). Primers were synthesized by Midland Certified Reagents (Midland, TX) and were purified by anion-exchange chromatography. Agarose gels and ethidium bromide were from Sigma. All other reagents were obtained from Perkin-Elmer. 3. Results a. Antigen Production and mRNA in Various Cell Lines Table 7 illustrates that a wide variety of human cancer cells from established lines express CFHφ tumor antigen as determined by presence of immunologically active antigen in the cell culture media and the appropriate mRNA within the cells. Myeloid lines, a human colon cancer line, and normal human epithelial keratinocytes (a primary culture, not an established cell line) are negative for antigen expression by both assay methods used. In contrast, many bladder, renal, cervical and prostate cancer cell lines produce this tumor antigen. Table 7 Production of huCFH by Various Cell Lines Cell Line Tumor Source sEIA RT-PCR (Media) RCC7860 Renal Cell Ca. Neg ++ ACHN rt Neg (+) Pastor + ++ 769P + ++ CAKI-1 Renal Clear Cell Neg (+) HL60 Myeloid Neg Neg LSI74T Colon AdenoCa. Neg Neg T24 TCC, Bladder 3+ ++ 5637 Primary Bladder CA Neg + RT4 Papillary Bladder CA 3+ ++ Cell Line Tumor Source sEIA RT-PCR (Media) J82 TCC, Bladder 3+ (+) 486P TCC, Bladder 3+ ++ HTB5 TCC, Bladder Neg + HTB9 TCC, Bladder 2+ ++ DU145 Prostate Ca. ND + PC3 Prostate Ca. ND + LNCaP Prostate Ca. ND (+) HeLaS3 Cervical AdenoCa. 4+ ++ HTB33 Cervical ND + C4I Cervical ND + DMEM/10%FBS Cell Culture Media (not Neg NA exposed to human cells) NHEK Normal human epithelial Neg Neg keratinocytes, NOT an established cell line X44.1 mouse hybridoma Neg Neg X52.1 Neg Neg X13.2 Neg Neg NA, not applicable; ND, not determined b. Analysis of RT-PCR Amplification Products Total RNA was set at 3 μg and 40 cycles of amplification were performed. Annealing temperatures were set at 50°C for 42M1040RT pairs, 56°C for 410M1040RT and 70°C for 2910M3610RT. The 42M1040RT product expressed with mRNA from cervical adenocarcinoma HeLaS3 cells is of the expected size for a human CFH-derived product and includes the 5' UTR or CFH, to which the 42M primer hybridizes. A second upstream primer, designated 410M (ACATGTAATGAGGGGTATCAA) (SEQ ID NO:20) and also derived from the huCFH sequence (GenBank Accession number Y00716), also yields a product of expected size and restriction map from HeLaS3 cells. In addition, RT-PCR of total RNA from TCC bladder cancer cell line HTB9 yields a cDNA of an appropriate size and restriction map using the 410M1040RT primer pair. The same primer pair yields no amplicon at the expected size for the colon adenocarcinoma line LS174T, myeloblastoma line HL-60 or normal human epithelial keratinocytes (NHEK). The experiments with preparations of HeLaS3 and HTB9 total RNA utilizing the 42M1040RT primer pair produced not only cDNA of the expected size, but also amplicons of unexpected sizes at 800 base pairs and, just above the limit of detectability, at 450, 480 and 1400 base pairs. In contrast, under the same conditions, the LS174T cell line produces only cDNA of incorrect sizes at 800, 450 and 420 base pairs, as well as an amplicon of 1200 base pairs which is at the limit of detectability. Alterations in the CFH gene product are not restricted to the 5' end of the molecule. Although amplification of the downstream portion of the HeLaS3 cDNA with CFH primers 2910M (GTCAGACAGTTATCAGTATGGAGAAGAAG) (SEQ ID NO: 18) and 361 ORT (CTGTTTGGCTGTCCACCTTAATGCTATG) (SEQ ID NO: 19) yielded amplicons of the expected size, the same primer pair yielded no amplicon with RNA from LS174T. Furthermore, both NHEK and HL-60 lines remained negative for the expression of CFHφ. Figure 5 shows the gel electrophoresis of amplification products resulting from RT-PCR performed with three primer sets derived from human complement Factor H (lanes 1 to 10 beginning at the left side of the gel with the left side set of numbers 1-4 on the Figure representing lanes 1-4, the middle set of numbers 1 -4 representing lanes 6-9 with lane 5 preceding, and the right side set of numbers 1 -4 representing lanes 11-14 with lane 10 preceding). Lane LHTB-9 product with primers 1040RT and 42M; Lane 2: HeLaS3 product with primers 1040RT and 42M; Lane 3: NHEK product with primers 1040RT and 42M; Lane 4: LS174T product with primers 1040RT and 42M; Lane 6: HTB-9 product with primers 1040RT and 410M; Lane 7: HeLaS3 product with primers 1040RT and 410M; Lane 8: NHEK product with primers 1040RT and 410M; Lane 9: LS174T product with primers 1040RT and 410M; Lane 11 : HTB-9 product with primers 361 ORT and 2910M; Lane 12: HeLaS3 product with primers 361 ORT and 2910M; Lane 13: NHEK product with primers 361 ORT and 2910M; Lane 14: LS174T product with primers 3610RT and 2910M; Lanes 5 and 10: DNA molecular weight markers. B. Method of Cloning cDNA Coding for CFHrp A cDNA of the appropriate size and restriction pattern for human CFH was prepared from mRNA isolated from the HeLaS3 human cervical adenocarcinoma cell line. This cDNA, prepared with primers 42M and 1040RT, was blunt-end cloned into pBluescript SK (Stratagene, La Jolla, CA). 1. Cloning and Sequencing a. PCR product, in a total volume of 73 μL, was purified using a Prep-A- Gene DNA purification kit (Stratagene). To the product in solution, 360 μL of the purification kit binding buffer and 20 μL of Prep-A-Gene DNA binding matrix were added. Purification was then performed according to the protocol described in the package insert. DNA concentration was estimated by running 2 μL of the purified material on a 2% agarose gel (Sigma) and comparing its intensity to the intensity of a standard (Sigma) of known concentration. b. To ensure that the PCR product had blunt ends, the ends were filled in using the PCR Polishing Kit (Stratagene). 1 μL of dNTP mix and 1 μL of Pfu Polymerase were added to 7 μL of purified PCR product to which had been added 1 μL of 10X Pfu Polymerase Buffer. The reaction mixture was incubated at 72°C for 30 minutes. Polished reactions were stored at -80°C until use. No further purification was required. c. In order to yield blunt ends, 20 μg of pBluescnpt SK- was digested with 30 units of Eco RV in a 100 μL reaction volume containing 6 mM Tris- HCI, pH 7.9, 6 mM MgCl 2 , 150 mM NaCI, and 1 mM DTT. The reaction mixture was incubated ovemight (16-18 hours) at 37°C. d. Phenol extraction of the digested plasmid was performed by adding an equal volume of Buffer-Saturated Phenol (Sigma) and vortexing to mix. The mixture was centrifuged at 5000 φm for 5 minutes to separate the phases. The upper aqueous phase was removed and transferred to a new tube. To the aqueous solution were added 50 μL phenol and 50 μL chloroform:isoarnyl alcohol (24: 1). The phases were mixed by vortexing and separated as above. The upper aqueous phase was removed and transferred to a new tube. This aqueous phase was again extracted with 100 ul of chloroform:isoamyl alcohol. The phases were separated as before, and the upper aqueous phase was removed and transferred to a fresh tube. e. In order to precipitate DNA from the aqueous solution, 0.5 volumes of 7.5 M ammonium acetate (pH 5.5) and 2 volumes of ethanol were added and incubated at -20°C for 1 hour. DNA was pelleted by centrifugation at 12,000xG for 20 minutes. The ethanol was carefully decanted. The DNA pellet was washed with 500 μL of 70% ethanol and then pelleted and supernatant decanted as above. The DNA pellet was allowed to air dry by inverting the tube until just dry by visual inspection and was then resuspended in water (Molecular Biology Grade, Sigma). f. 10 μg of digested pBluescript DNA was dephosphorylated with 500 units of Calf Intestinal Alkaline Phosphatase by incubating at 37°C for 15 minutes in a total volume of 100 μL, containing 0.05 M Tris-HCI, pH 9.3, 1 mM MgCl 2 , 0.1 mM ZnCl 2 and 1 mM Spermidine. Another 500 units of alkaline phosphatase was added and reaction was incubated at 56°C for 45 minutes. The reaction mixture was then incubated at 65°C for 1 hour to inactivate the alkaline phosphatase. The reaction mixture was extracted with phenol and chloroform and then precipitated with ethanol, as described in step (d). g. Ligation of vector and insert was performed using the two-step Hgation procedure described by S. Damak and D.W. Bullock (BioTechniques 75(3):448-452, 1993). In the first step of the ligation process, vector and insert were mixed together at vectoπinsert ratio of 1:4. 150 nmol of polished insert was mixed with 36 nmol of vector in a 10 μL volume containing 0.05 M Tris, 0.025 M MgCl 2 , 0.5 mM ATP, 1 mM DTT, 1 unit of T4 DNA Ligase and 10 units of EcoRV. The reaction was incubated at room temperature for 2 hours For the second part of the ligation process, 190 μL of a solution containing 0.05 M Tris, 0.025 M MgCl 2 , 0.5 mM ATP, 1 mM DTT, and 19 units of T4 DNA Ligase was added to the reaction mixture from the first step and incubated at room temperature ovemight (16-18 hours). h. Transformation of competent E. coli was initiated by first thawing frozen competent DH5 (Stratagene) cells in an ice water bath. 1.7 mL microfuge tubes were labelled and placed on ice. When thawed, 50 μL of the competent cells was pipetted into the labelled tubes. 5 μL of the ligation reaction mixture was added to the tube containing the competent cells. The tube was flicked briefly to mix and incubated on ice for 30 minutes. This tube containing the competent cell/ligation mixture was then incubated at 37°C for 20 seconds. A 0.95 mL volume of LB broth (Sigma) was added and tubes were incubated at 37°C for 1 hour with shaking at 225 φm. 200 μL of cell mixture was spread, using sterile spreaders, onto the surface of an LB agar plate containing 50 μg/ml Ampicillin, 25 μg/ml X-Gal and 60 μg/ml IPTG (all reagents from Sigma). Plates were covered and incubated at 37°C overnight (16-18 hours). White colonies were selected and screened by purifying the plasmid DNA and then performing restriction analysis. All DNA sequencing was performed on ABI Prism sequencers (Perkin- Elmer, Foster City, CA) in the laboratory of Dr. Leroy Hood, Center for Molecular Biotechnology, University of Washington, Seattle, WA. Results Table 8 is a tabulation of observed differences between the clone sequences (for DNA encoding complement Factor H-related protein) and the published sequence for DNA encoding human complement Factor H (CFH). Table 8 Apparent Differences Between Clone and Human CFH Sequences DNA Mature Amino Clone RNA Position DNA Protein Acid Source Number Change Position Change Number pRBB9FH410#2.1 HTB9 672 AtoG 181 StoG pRBS3FH2910#2.1 Hela S3 3096 GtoT 989 VtoL pRBS3FH2910#3.1 Hela S3 3046 TtoC 972 LtoP 3094 AtoG 988 DtoV 3096 GtoT 989 VtoL 3142 AtoG 1004 YtoC 3144 AtoC 1005 KtoQ pRBS3FH2910#4.1 Hela S3 3053 AtoG 974 No Change 3096 GtoT 989 VtoL 3115 AtoC 995 QtoP pZS3FH2576#3 Hela S3 2621 AtoG 830 No Change 2746 GtoT 872 Stol pZS3FH2576#l/ll Hela S3 2746 GtoT 872 Stol Figure 6A shows a partial DNA sequence from clone pRBB9FH410 and Figure 6B the corresponding amino acid sequence, as compared to the DNA and amino acid sequences for human CFH. Figure 7A shows three partial DNA sequences from clone pRBS3FH2910 and Figure 7B the corresponding amino acid sequences, as compared to the DNA and amino acid sequences for human CFH. Figure 8A shows two partial DNA sequences from clone pZS3FH2576 and Figure 8B the corresponding amino acid sequences, as compared to the DNA and amino acid sequences for human CFH. 2. Generation and Use of Riboprobes a. A riboprobe for use in in situ hybridization was prepared from the same clone. Restriction digestion of the clone was performed with Ava II (New England Biolabs, Beverly, MA) following the manufacturer's instructions. The resulting cD A corresponded to map positions 457- 1057 of huCFH (GenBank sequence number YM00716). The cDNA product was purified by electrophoresis on a 1% agarose TAE gel (Sigma). A digoxin-labelled ribonucleic acid antisense probe was prepared with T7 RNA polymerase, using a commercial kit (Boehringer- Mannheim, Indianapolis, IN) and following the instructions in the package insert. Unlabelled RNA for probe competition was synthesized in a similar manner using a kit from Ambion (Austin, TX) following the manufacturer's instructions. b. The resulting Riboprobes were purified by precipitation with an equal volume of absolute ethanol and were then redissolved in ribonuclease- free water to final concentrations of either 101 nM for the digoxigenin- labelled probe or 10 uM for the unlabelled probe. c. Pathology tissue specimens were prepared for staining by snap freezing in liquid nitrogen. The frozen pellets were sectioned on a cryostat microtome (Bartles & Stout) and then fixed in (-20°C) acetone (Sigma) for 15 minutes. Fixed sections were placed on a slide and kept wet in APK buffer (Ventana, Tucson, AZ) until the hybridization process was begun. d. For tissue staining, 1 μL of stock riboprobe was diluted into 500 μL of hybridization solution, consisting of 2X Denhardt's solution supplemented with 60% (w/v) formamide, 12.5% dextran sulfate, 10 mM Tris, 1 mM EDTA, 1 mM DTT, 375 mM NaCI, 0.3% Triton X100, and containing 2 mg tRNA (all reagents from Sigma). Final concentration of the digoxigenin-labelled RNA probe was 0.3 nM. e. Staining was performed on an ES Genii slide processor (Ventana, Tucson, AZ), using reagent packs and buffers from the manufacturer, with detection by HRP-conjugated anti-mouse antibody. Hybridization solution containing the riboprobe was applied manually and processed wet. Following denaturation for 2 minutes at 65°C, hybridization was carried out at 40°C for 120 minutes. Three washes were performed at 55°C sequentially with IX SSC (150 mM sodium chloride, 15 mM sodium citrate, pH 7.0-7.5) 0.5X SSC and 0.1X SSC. Slides were then reacted with anti-digoxigenin antibody following the manufacturer's instructions (Boehringer Mannheim). A specificity control was performed by application of riboprobe stock containing a 100-fold excess of riboprobe that had not been labeled with digoxigenin. Results The target tissues subjected to staining with the riboprobe were serial sections from normal and cancerous human bladder (transitional cells) and from normal and cancerous human prostate. All tissue sections, both normal and cancerous, were from a single bladder or a single prostate. Specificity of tissues staining was established by competition of digoxigenin labelled probe binding with a 100-fold excess of unlabelled probe. Only sections from TCC+ bladder cancer stained with the HeLaS3 generated probe sequence. EXAMPLE VII INHIBITION OF ANTIGEN BIOLOGICAL ACTIVITY BY MABS A. In Vitro Protection of C3b bv Anti-CFH Related Protein MAbs As shown in Example III.F the complement Factor H-related activity of antigen can be mimicked by complement Factor H itself. For experimental clarity, therefore, Factor H and Factor I were used to degrade C3b and illustrate the protective actions of anti-Factor H-related protein MAbs. Reactions were performed by incubating 1 μg of Factor H with either 15 or 30 μg of each of three MOF MAbs (X52.1; X87.2; XI 3.2), in 20 μL of phosphate- buffered saline for 30 minutes at 37°C, followed by the addition of 7.5 μg of C3b and 5 μg of Factor I into each reaction tube (final reaction volume 32.5 μL). (C3b was generated from C3 by the method of Pangburn, M.K., and Mueller-Eberhard, H.J., Biochemistry 22:178-185, 1983.) The mixture was then incubated on a rotator at 37°C for 1 hour. Results were determined by SDS-PAGE of the reaction mixtures under reducing conditions (50 mM DTT) on a 4-12% gradient gel (Novex, San Diego, CA). (Unless specified otherwise, all reagents are from Sigma, St. Louis; MO.) The gel was scanned with a GelDoc scanner (BioRad, Hercules, CA). The intensities of the bands measured in this way were converted to percentage of C3b remaining. The control lane containing the reaction mixture in the absence of MAb was used to represent 100 percent degradation, while the lane containing the reaction mixture with no Factor H was used to represent 0 percent degradation. Results The results derived from scanning the gel are summarized in Table 8. These results demonstrate that MAbs X13.2 and X87.2 are able to block the Factor H- related protein antigen-mediated degradation of complement component C3b in vitro. B. Activation of Cell Lysis by the Alternate Complement Pathway bv MAbs Specific for the Antigen Standard guinea pig complement was treated with 5 M EGTA to chelate calcium. Then 5 mM MgCl 2 , which is required for the activity of the alternative complement pathway (ACP), was added. The mixture was incubated for 20 minutes at 37°C, then added to 7 x IO 9 rabbit red blood cells (RBC) and further incubated at 37°C. After 45 minutes and again after 117 minutes, hemolysis was determined by measuring the A 450 and comparing the values to those determined for control reactions, which had received either no complement or no MAb. Measurements were performed on a Dynatech (Chantilly, VA) MR5000 96-well microplate reader. Hemolysis was determined in the absence or presence of MAb X52.1. When included, the MAb was used at a concentration in the reaction mixture of 10 nM or 30 nM (Figure 3). All reagents and materials were purchased from Sigma. A second experiment was performed under the same conditions as the RBC lysis, but the target cells were HL-60 (1 x IO 8 cells), a human myeloid cell line. MAb concentration was set to 10 nM, and lysis was read after 120 minutes as described above (Figure 4). Results Although MAb X52.1 was not effective in the format used in Example VILA above, perhaps due to a lower affinity for Factor H, it was highly effective in promoting lysis of RBCs or HL-60 cells. RBC lysis was dependent on the concentration of MAb and the duration of incubation. Since X52.1 does not bind to RBCs or HL-60 cells, and since HL-60 cells do not produce Factor H, the mechanism of action of cell lysis must be the binding of X52.1 to the Factor H in the added guinea pig complement and resulting inhibition of the C3 stabilization activity of the Factor H. In summary, MAb X52.1, which specifically binds antigen and CFH, but does not bind red blood cells, Factor I or C3b, can promote the ACP-mediated lysis of RBCs. The concentrations required to do this are achievable physiologically (10 nM, or approximately 1.5 μg/mL). Table 9 Inhibition of C3b Degradation in the Presence of Anti- Antigen MAbs Sample Quantity of Sample C3b Remaining (Percent) Control, No MAb Standard 0 X52.1 15 μg 0 X52.1 30 μg 0 X87.2 15 μg 24.7 X87.2 30 μg 54.9 X13.2 15 μg 62.0 X13.2 30 μg 54.0 Control, No Factor H Standard 100 All publications and patent applications mentioned in this specification are herein incoφorated by reference to the same extent as if each individual publication or patent application was specifically and individually incoφorated by reference. From the foregoing, it will be appreciated that, although specific embodiments of the invention have been described herein for puφoses of illustration, various modifications may be made without deviating from the spirit and scope of the invention. SEQUENCE LISTING (1) GENERAL INFORMATION: (l) APPLICANT: Kmders, Robert J. Enfield, David L. Hass, G. Michael (ii) TITLE OF INVENTION: METHODS AND COMPOSITIONS FOR SCREENING FOR OR MODULATING A TUMOR ASSOCIATED ANTIGEN (in) NUMBER OF SEQUENCES: 38 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: SEED and BERRY LLP (B) STREET: 6300 Columbia Center, 701 Fifth Avenue (C) CITY: Seattle (D) STATE: Washington (E) COUNTRY: USA (F) ZIP: 98104 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.30 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: US (B) FILING DATE: 09-APR-1997 (C) CLASSIFICATION: (vm) ATTORNEY/AGENT INFORMATION: (A) NAME: Sharkey, Richard G. (B) REGISTRATION NUMBER: 32,629 (C) REFERENCE/DOCKET NUMBER: 130001.404Pc (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (206) 622-4900 (B) TELEFAX: (206) 682-6031 (2) INFORMATION FOR SEQ ID NO:l: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: Glu Asp Cys Asn Xaa Leu Pro Pro Arg Xaa Asn Thr 1 5 10 (2) INFORMATION FOR SEQ ID NO: 2: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Gly Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr Tyr Xaa Tyr Tyr Xaa 1 5 10 15 Asp (2) INFORMATION FOR SEQ ID NO:3: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr Tyr Ser Xaa Tyr Xaa 1 5 10 15 Asp Glu Xaa Phe Xaa Xaa Xaa Ser 20 (2) INFORMATION FOR SEQ ID NO: : (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: am o acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (Xl) SEQUENCE DESCRIPTION: SEQ ID NO: : Ser Ser Gin Glu Ser Tyr Ala His Gly Thr Lys 1 5 10 (2) INFORMATION FOR SEQ ID NO: 5: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: Glu Asp Cys Asn Glu Leu Pro Pro Xaa Arg Asn Thr Glu lie Leu Xaa 1 5 10 15 Gly Ser Trp Asp 20 (2) INFORMATION FOR SEQ ID NO: 6: (ι) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: Arg Pro Tyr Phe Pro Val Val Ala Val Gly Lys Tyr Tyr Ser Tyr Tyr 1 5 10 15 Xaa Asp Glu His Phe Glu Xaa Pro 20 (2) INFORMATION FOR SEQ ID NO: 7: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: Ser Leu Gly Asn Val lie Met Val Gly Arg Lys Gly Glu Trp Val Ala 1 5 10 15 Leu Asn Pro Leu Arg Lys 20 (2) INFORMATION FOR SEQ ID NO: 8: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr 1 5 10 (2) INFORMATION FOR SEQ ID NO: 9: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: TCGTTCATTC TCCTTAT 17 (2) INFORMATION FOR SEQ ID NO: 10: (ι) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: GCTGGTAAAT GTCCTCT 17 (2) INFORMATION FOR SEQ ID NO: 11: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: ATGTAATGAG GGGTATC 17 (2) INFORMATION FOR SEQ ID NO: 12: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 amino acids (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12: Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr Tyr Ser Tyr Tyr Cys 1 5 10 15 Asp (2) INFORMATION FOR SEQ ID NO: 13: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 ammo acids (C) STRANDEDNESS: (D) TOPOLOGY: linear (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr Tyr Ser Tyr Tyr Cys 1 5 10 15 Asp Glu His Phe Glu Thr Pro Ser 20 (2) INFORMATION FOR SEQ ID NO: 14: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 ammo acids (C) STRANDEDNESS: (D) TOPOLOGY: linear ( xi ) SEQUENCE DESCRI PTI ON : SEQ I D NO : 14 : Gl u Asp Cys Asn Glu Leu Pro Pro Arg Arg Asn Thr Gl u He Leu Thr 1 5 10 15 Gly Ser Trp Ser Asp 20 (2) INFORMATION FOR SEQ ID NO: 15: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 23 ammo acids (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15: Arg Pro Tyr Phe Pro Val Ala Val Gly Lys Tyr Tyr Ser Tyr Tyr Cys 1 5 10 15 Asp Glu His Phe Glu Thr Pro 20 (2) INFORMATION FOR SEQ ID NO: 16: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 ammo acids (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: Ser Leu Gly Asn Val He Met Val Cys Arg Lys Gly Glu Trp Val Ala 1 5 10 15 Leu Asn Pro Leu Arg Lys 20 (2) INFORMATION FOR SEQ ID NO: 17: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: TCTGGATAAT CACAAGGTTT C 21 (2) INFORMATION FOR SEQ ID NO: 18: (ι) SEQUENCE CHARACTERISTICS: (A) LENGTH: 29 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (ϋ) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: GTCAGACAGT TATCAGTATG GAGAAGAAG 29 (2) INFORMATION FOR SEQ ID NO: 19: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 28 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19: CTGTTTGGCT GTCCACCTTA ATGCTATG 28 (2) INFORMATION FOR SEQ ID NO:20: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: ACATGTAATG AGGGGTATCA A 21 (2) INFORMATION FOR SEQ ID NO: 21: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 649 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: ACATGTAATG AGGGGTATCA ATTGCTAGGT GAGATTAATT ACCGTGAATG TGACACAGAT 60 GGATGGACCA ATGATATTCC TATATGTGAA GTTGTGAAGT GTTTACCAGT GACAGCACCA 120 GAGAATGGAA AAATTGTCAG TAGTGCAATG GAACCAGATC GGGAATACCA TTTTGGACAA 180 GCAGTACGGT TTGTATGTAA CTCAGGCTAC AAGATTGAAG GAGATGAAGA AATGCATTGT 240 TCAGACGATG GTTTTTGGAG TAAAGAGAAA CCAAAGTGTG TGGAAATTTC ATGCAAATCC 300 CCAGATGTTA TAAATGGATC TCCTATATCT CAGAAGATTA TTTATAAGGA GAATGAACGA 360 TTTCAATATA AATGTAACAT GGGTTATGAA TACAGTGAAA GAGGAGATGC TGTATGCACT 420 GAATCTGGAT GGCGTCCGTT GCCTTCATGT GAAGAAAAAT CATGTGATAA TCCTTATATT 480 CCAAATGGTG ACTACTCACC TTTAAGGATT AAACACAGAA CTGGAGATGA AATCACGTAC 540 CAGTGTAGAA ATGGTTTTTA TCCTGCAACC CGGGGAAATA CAGCCAAATG CACAAGTACT 600 GGCTGGATAC CTGCTCCGAG ATGTACCTTG AAACCTTGTG ATTATCCAG 649 (2) INFORMATION FOR SEQ ID NO: 22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 581 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: ACATGTAATG AGGGGTATCA ATTGCTAGGT GAGATTAATT ACCGTGAATG TGACACAGAT 60 GGATGGACCA ATGATATTCC TATATGTGAA GTTGTGAAGT GTTTACCAGT GACAGCACCA 120 GAGAATGGAA AAATTGTCAG TAGTGCAATG GAACCAGATC GGGAATACCA TTTTGGACAA 180 GCAGTACGGT TTGTATGTAA CTCAGGCTAC AAGATTGAAG GAGATGAAGA AATGCATTGT 240 TCAGACGATG GTTTTTGGGG TAAAGAGAAA CCAAAGTGTG TGGAAATTTC ATGCAAATCC 300 CCAGATGTTA TAAATGGATC TCCTATATCT CAGAAGATTA TTTATAAGGA GAATGAACGA 360 TTTCAATATA AATGTAACAT GGGTTATGAA TACAGTGAAA GAGGAGATGC TGTATGCACT 420 GAATCTGGAT GGCGTCCGTT GCCTTCATGT GAAGAAAAAT CATGTGATAA TCCTTATATT 480 CCAAATGGTG ACTACTCACC TTTAAGGATT AAACACAGAA CTGGAGATGA AATCACGTAC 540 CAGTGTAGAA ATGGTTTTTA TCCTGCAACC CGGGGAAATA C 581 (2) INFORMATION FOR SEQ ID NO:23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 240 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Phe Thr Leu Thr Gly Gly Asn Val Phe Glu Tyr Gly Val Lys Ala Val 1 5 10 15 Tyr Thr Cys Asn Glu Gly Tyr Gin Leu Leu Gly Glu He Asn Tyr Arg 20 25 30 Glu Cys Asp Thr Asp Gly Trp Thr Asn Asp He Pro He Cys Glu Val 35 40 45 Val Lys Cys Leu Pro Val Thr Ala Pro Glu Asn Gly Lys He Val Ser 50 55 60 Ser Ala Met Glu Pro Asp Arg Glu Tyr His Phe Gly Gin Ala Val Arg 65 70 75 80 Phe Val Cys Asn Ser Gly Tyr Lys He Glu Gly Asp Glu Glu Met His 85 90 95 Cys Ser Asp Asp Gly Phe Trp Ser Lys Glu Lys Pro Lys Cys Val Glu 100 105 110 He Ser Cys Lys Ser Pro Asp Val He Asn Gly Ser Pro He Ser Gin 115 120 125 Lys He He Tyr Lys Glu Asn Glu Arg Phe Gin Tyr Lys Cys Asn Met 130 135 140 Gly Tyr Glu Tyr Ser Glu Arg Gly Asp Ala Val Cys Thr Glu Ser Gly 145 150 155 160 Trp Arg Pro Leu Pro Ser Cys Glu Glu Lys Ser Cys Asp Asn Pro Tyr 165 170 175 He Pro Asn Gly Asp Tyr Ser Pro Leu Arg He Lys His Arg Thr Gly 180 165 190 Asp Glu He Thr Tyr Gin Cys Arg Asn Gly Phe Tyr Pro Ala Thr Arg 195 200 205 Gly Asn Thr Ala Lys Cys Thr Ser Thr Gly Trp He Pro Ala Pro Arg 210 215 220 Cys Thr Leu Lys Pro Cys Asp Tyr Pro Asp He Lys His Gly Gly Leu 225 230 235 240 (2) INFORMATION FOR SEQ ID NO:24: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 216 am no acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: Thr Cys Asn Glu Gly Tyr Gin Leu Leu Gly Glu He Asn Tyr Arg Glu 1 5 10 15 Cys Asp Thr Asp Gly Trp Thr Asn Asp He Pro He Cys Glu Val Val 20 25 30 Lys Cys Leu Pro Val Thr Ala Pro Glu Asn Gly Lys He Val Ser Ser 35 40 45 Ala Met Glu Pro Asp Arg Glu Tyr His Phe Gly Gin Ala Val Arg Phe 50 55 60 Val Cys Asn Ser Gly Tyr Lys He Glu Gly Asp Glu Giu Met His Cys 65 70 75 80 Ser Asp Asp Gly Phe Trp Gly Lys Glu Lys Pro Lys Cys Val Glu He 85 90 95 Ser Cys Lys Ser Pro Asp Val He Asn Gly Ser Pro He Ser Gin Lys 100 105 110 He He Tyr Lys Glu Asn Glu Arg Phe Gin Tyr Lys Cys Asn Met Gly 115 120 125 Tyr Glu Tyr Ser Glu Arg Gly Asp Ala Val Cys Thr Glu Ser Gly Trp 130 135 140 Arg Pro Leu Pro Ser Cys Glu Glu Lys Ser Cys Asp Asn Pro Tyr He 145 150 155 160 Pro Asn Gly Asp Tyr Ser Pro Leu Arg He Lys His Arg Thr Gly Asp 165 170 175 Glu He Thr Tyr Gin Cys Arg Asn Gly Phe Tyr Pro Ala Thr Arg Gly 180 185 190 Asn Thr Ala Lys Cys Thr Ser Thr Gly Trp He Pro Ala Pro Arg Cys 195 200 205 Thr Leu Lys Pro Cys Asp Tyr Pro 210 215 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 767 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: CATGGTGTTG TAGCTCACAT GTCAGACAGT TATCAGTATG GAGAAGAAGT TACGTACAAA 60 TGTTTTGAAG GTTTTGGAAT TGATGGGCCT GCAATTGCAA AATGCTTAGG AGAAAAATGG 120 TCTCACCCTC CATCATGCAT AAAAACAGAT TGTCTCAGTT TACCTAGCTT TGAAAATGCC 180 ATACCCATGG GAGAGAAGAA GGATGTGTAT AAGGCGGGTG AGCAAGTGAC TTACACTTGT 240 GCAACATATT ACAAAATGGA TGGAGCCAGT AATGTAACAT GCATTAATAG CAGATGGACA 300 GGAAGGCCAA CATGCAGAGA CACCTCCTGT GTGAATCCGC CCACAGTACA AAATGCTTAT 360 ATAGTGTCGA GACAGATGAG TAAATATCCA TCTGGTGAGA GAGTACGTTA TCAATGTAGG 420 AGCCCTTATG AAATGTTTGG GGATGAAGAA GTGATGTGTT TAAATGGAAA CTGGACGGAA 480 CCACCTCAAT GCAAAGATTC TACAGGAAAA TGTGGGCCCC CTCCACCTAT TGACAATGGG 540 GACATTACTT CATTCCCGTT GTCAGTATAT GCTCCAGCTT CATCAGTTGA GTACCAATGC 600 CAGAACTTGT ATCAACTTGA GGGTAACAAG CGAATAACAT GTAGAAATGG ACAATGGTCA 660 GAACCACCAA AATGCTTACA TCCGTGTGTA ATATCCCGAG AAATTATGGA AAATTATAAC 720 ATAGCATTAA GGTGGACAGC CAAACAGAAG CTTTATTCGA GAACAGG 767 (2) INFORMATION FOR SEQ ID NO: 26: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 532 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: GTCAGACAGT TATCAGTATG GAGAAGAAGT TACGTACAAA TGTTTTGAAG GTTTTGGAAT 60 TGATGGGCCT GCAATTGCAA AATGCTTAGG AGAAAAATGG TCTCACCCTC CATCATGCAT 120 AAAAACAGAT TGTCTCAGTT TACCTAGCTT TGAAAATGCC ATACCCATGG GAGAGAAGAA 180 GGATTTGTAT AAGGCGGGTG AGCAAGTGAC TTACACTTGT GCAACATATT ACAAAATGGA 240 TGGAGCCAGT AATGTAACAT GCATTAATAG CAGATGGACA GGAAGGCCAA CATGCAGAGA 300 CACCTCCTGT GTGAATCCGC CCACAGTACA AAATGCTTAT ATAGTGTCGA GACAGATGAG 360 TAAATATCCA TCTGGTGAGA GAGTACGTTA TCAATGTAGG AGCCCTTATG AAATGTTTGG 420 GGATGAAGAA GTGATGTGTT TAAATGGAAA CTGGACGGAA CCACCTCAAT GCAAAGATTC 480 TACAGGAAAA TGTGGGCCCC CTCCACCTAT TGACAATGGG GACATTACTT CA 532 (2) INFORMATION FOR SEQ ID NO: 27: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 688 base pairs (B) TYPE: nucleic ac d (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: GTCAGACAGT TATCAGTATG GAGAAGAAGT TACGTACAAA TGTTTTGAAG GTTTTGGAAT 60 TGATGGGCCT GCAATTGCAA AATGCTTAGG AGAAAAATGG TCTCACCCTC CATCATGCAT 120 AAAAACAGAT TGTCCCAGTT TACCTAGCTT TGAAAATGCC ATACCCATGG GAGAGAAGAA 180 GGTTTTGTAT AAGGCGGGTG AGCAAGTGAC TTACACTTGT GCAACATATT GCCAAATGGA 240 TGGAGCCAGT AATGTAACAT GCATTAATAG CAGATGGACA GGAAGGCCAA CATGCAGAGA 300 CACCTCCTGT GTGAATCCGC CCACAGTACA AAATGCTTAT ATAGTGTCGA GACAGATGAG 360 TAAATATCCA TCTGGTGAGA GAGTACGTTA TCAATGTAGG AGCCCTTATG AAATGTTTGG 420 GGATGAAGAA GTGATGTGTT TAAATGGAAA CTGGACGGAA CCACCTCAAT GCAAAGATTC 480 TACAGGAAAA TGTGGGCCCC CTCCACCTAT TGACAATGGG GACATTACTT CATTCCCGTT 540 GTCAGTATAT GCTCCAGCTT CATCAGTTGA GTACCAATGC CAGAACTTGT ATCAACTTGA 600 GGGTAACAAG CGAATAACAT GTAGAAATGG ACAATGGTCA GAACCACCAA AATGCTTACA 660 TCCGTGTGTA ATATCCCGAG AAATTATG 688 (2) INFORMATION FOR SEQ ID NO:28: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 590 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: GTTTGCCTAG CTTTGAAAAT GCCATACCCA TGGGAGAGAA GAAGGATTTG TATAAGGCGG 60 GTGAGCCAGT GACTTACACT TGTGCAACAT ATTACAAAAT GGATGGAGCC AGTAATGTAA 120 CATGCATTAA TAGCAGATGG ACAGGAAGGC CAACATGCAG AGACACCTCC TGTGTGAATC 180 CGCCCACAGT ACAAAATGCT TATATAGTGT CGAGACAGAT GAGTAAATAT CCATCTGGTG 240 AGAGAGTACG TTATCAATGT AGGAGCCCTT ATGAAATGTT TGGGGATGAA GAAGTGATGT 300 GTTTAAATGG AAACTGGACG GAACCACCTC AATGCAAAGA TTCTACAGGA AAATGTGGGC 360 CCCCTCCACC TATTGACAAT GGGGACATTA CTTCATTCCC GTTGTCAGTA TATGCTCCAG 420 CTTCATCAGT TGAGTACCAA TGCCAGAACT TGTATCAACT TGAGGGTAAC AAGCGAATAA 480 CATGTAGAAA TGGACAATGG TCAGAACCAC CAAAATGCTT ACATCCGTGT GTAATATCCC 540 GAGAAATT T GGAAAATTAT AACATAGCAT TAAGGTGGAC AGCCAAACAG 590 (2) INFORMATION FOR SEQ ID NO:29: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 290 ammo acids (C) STRANDEDNESS: ( D ) TOPOLOGY : linear ( xi ) SEQUENCE DESCRI PTION : SEQ I D NO : 29 : Gl y Phe Arg He Ser Glu Glu Asn Glu Thr Thr Cys Tyr Met Gl y Lys 1 5 10 15 Trp Ser Ser Pro Pro Gin Cys Glu Gly Leu Pro Cys Lys Ser Pro Pro 20 25 30 Glu He Ser His Gly Val Val Ala His Met Ser Asp Ser Tyr Gin Tyr 35 40 45 Gly Glu Glu Val Thr Tyr Lys Cys Phe Glu Gly Phe Gly He Asp Gly 50 55 60 Pro Ala He Ala Lys Cys Leu Gly Glu Lys Trp Ser His Pro Pro Ser 65 70 75 80 Cys He Lys Thr Asp Cys Leu Ser Leu Pro Ser Phe Glu Asn Ala He 85 90 95 Pro Met Gly Glu Lys Lys Asp Val Tyr Lys Ala Gly Glu Gin Val Thr 100 105 110 Tyr Thr Cys Ala Thr Tyr Tyr Lys Met Asp Gly Ala Ser Asn Val Thr 115 120 125 Cys He Asn Ser Arg Trp Thr Gly Arg Pro Thr Cys Arg Asp Thr Ser 130 135 140 Cys Val Asn Pro Pro Thr Val Gin Asn Ala Tyr He Val Ser Arg Gin 145 150 155 160 Met Ser Lys Tyr Pro Ser Gly Glu Arg Val Arg Tyr Gin Cys Arg Ser 165 170 175 Pro Tyr Glu Met Phe Gly Asp Glu Glu Val Met Cys Leu Asn Gly Asn 180 185 190 Trp Thr Glu Pro Pro Gin Cys Lys Asp Ser Thr Gly Lys Cys Gly Pro 195 200 205 Pro Pro Pro He Asp Asn Gly Asp He Thr Ser Phe Pro Leu Ser Val 210 215 220 Tyr Ala Pro Ala Ser Ser Val Glu Tyr Gin Cys Gin Asn Leu Tyr Gin 225 230 235 240 Leu Glu Gly Asn Lys Arg He Thr Cys Arg Asn Gly Gin Trp Ser Glu 245 250 255 Pro Pro Lys Cys Leu His Pro Cys Val He Ser Arg Glu He Met Glu 260 265 270 Asn Tyr Asn He Ala Leu Arg Trp Thr Ala Lys Gin Lys Leu Tyr Ser 275 280 285 Arg Thr 290 (2) INFORMATION FOR SEQ ID NO: 30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30: Ser Asp Ser Tyr Gin Tyr Gly Glu Glu Val Thr Tyr Lys Cys Phe Glu 1 5 10 15 Gly Phe Gly He Asp Gly Pro Ala He Ala Lys Cys Leu Gly Glu Lys 20 25 30 Trp Ser His Pro Pro Ser Cys He Lys Thr Asp Cys Leu Ser Leu Pro 35 40 45 Ser Phe Glu Asn Ala He Pro Met Gly Glu Lys Lys Asp Leu Tyr Lys 50 55 60 Ala Gly Glu Gin Val Thr Tyr Thr Cys Ala Thr Tyr Tyr Lys Met Asp 65 70 75 80 Gly Ala Ser Asn Val Thr Cys He Asn Ser Arg Trp Thr Gly Arg Pro 85 90 95 Thr Cys Arg Asp Thr Ser Cys Val Asn Pro Pro Thr Val Gin Asn Ala 100 105 110 Tyr He Val Ser Arg Gin Met Ser Lys Tyr Pro Ser Gly Glu Arg Val 115 120 125 Arg Tyr Gin Cys Arg Ser Pro Tyr Glu Met Phe Gly Asp Glu Glu Val 130 135 140 Met Cys Leu Asn Gly Asn Trp Thr Glu Pro Pro Gin Cys Lys Asp Ser 145 150 155 160 Thr Gly Lys Cys Gly Pro Pro Pro Pro He Asp Asn Gly Asp He Thr 165 170 175 Ser (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 229 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Ser Asp Ser Tyr Gin Tyr Gly Glu Glu Val Thr Tyr Lys Cys Phe Glu 1 5 10 15 Gly Phe Gly He Asp Gly Pro Ala He Ala Lys Cys Leu Gly Glu Lys 20 25 30 Trp Ser His Pro Pro Ser Cys He Lys Thr Asp Cys Pro Ser Leu Pro 35 40 45 Ser Phe Glu Asn Ala He Pro Met Gly Glu Lys Lys Val Leu Tyr Lys 50 55 60 Ala Gly Glu Gin Val Thr Tyr Thr Cys Ala Thr Tyr Cys Gin Met Asp 65 70 75 80 Gly Ala Ser Asn Val Thr Cys He Asn Ser Arg Trp Thr Gly Arg Pro 85 90 95 Thr Cys Arg Asp Thr Ser Cys Val Asn Pro Pro Thr Val Gin Asn Ala 100 105 110 Tyr He Val Ser Arg Gin Met Ser Lys Tyr Pro Ser Gly Glu Arg Val 115 120 125 Arg Tyr Gin Cys Arg Ser Pro Tyr Glu Met Phe Gly Asp Glu Glu Val 130 135 140 Met Cys Leu Asn Gly Asn Trp Thr Glu Pro Pro Gin Cys Lys Asp Ser 145 150 155 160 Thr Gly Lys Cys Gly Pro Pro Pro Pro He Asp Asn Gly Asp He Thr 165 170 175 Ser Phe Pro Leu Ser Val Tyr Ala Pro Ala Ser Ser Val Glu Tyr Gin 180 185 190 Cys Gin Asn Leu Tyr Gin Leu Glu Gly Asn Lys Arg He Thr Cys Arg 195 200 205 Asn Gly Gin Trp Ser Glu Pro Pro Lys Cys Leu His Pro Cys Val He 210 215 220 Ser Arg Glu He Met 225 (2) INFORMATION FOR SEQ ID NO: 32: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 196 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 32: Leu Pro Ser Phe Glu Asn Ala He Pro Met Gly Glu Lys Lys Asp Leu 1 5 10 15 Tyr Lys Ala Gly Glu Pro Val Thr Tyr Thr Cys Ala Thr Tyr Tyr Lys 20 25 30 Met Asp Gly Ala Ser Asn Val Thr Cys He Asn Ser Arg Trp Thr Gly 35 40 45 Arg Pro Thr Cys Arg Asp Thr Ser Cys Val Asn Pro Pro Thr Val Gin 50 55 60 Asn Ala Tyr He Val Ser Arg Gin Met Ser Lys Tyr Pro Ser Gly Glu 65 70 75 80 Arg Val Arg Tyr Gin Cys Arg Ser Pro Tyr Glu Met Phe Gly Asp Glu 85 90 95 Glu Val Met Cys Leu Asn Gly Asn Trp Thr Glu Pro Pro Gin Cys Lys 100 105 110 Asp Ser Thr Gly Lys Cys Gly Pro Pro Pro Pro He Asp Asn Gly Asp 115 120 125 He Thr Ser Phe Pro Leu Ser Val Tyr Ala Pro Ala Ser Ser Val Glu 130 135 140 Tyr Gin Cys Gin Asn Leu Tyr Gin Leu Glu Gly Asn Lys Arg He Thr 145 150 155 160 Cys Arg Asn Gly Gin Trp Ser Glu Pro Pro Lys Cys Leu His Pro Cys 165 170 175 Val He Ser Arg Glu He Met Glu Asn Tyr Asn He Ala Leu Arg Trp 180 185 190 Thr Ala Lys Gin 195 (2) INFORMATION FOR SEQ ID NO: 33: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 472 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33: CACAATATGA CAACCACACT GAATTATCGG GATGGAGAAA AAGTATCTGT TCTTTGCCAA 60 GAAAATTATC TAATTCAGGA AGGAGAAGAA ATTACATGCA AAGATGGAAG ATGGCAGTCA 120 ATACCACTCT GTGTTGAAAA AATTCCATGT TCACAACCAC CTCAGATAGA ACACGGAACC 180 ATTAATTCAT CCAGGTCTTC ACAAGAAAGT TATGCACATG GGACTAAATT GAGTTATACT 240 TGTGAGGGTG GTTTCAGGAT ATCTGAAGAA AATGAAACAA CATGCTACAT GGGAAAATGG 300 AGTTCTCCAC CTCAGTGTGA AGGCCTTCCT TGTAAATCTC CACCTGAGAT TTCTCATGGT 360 GTTGTAGCTC ACATGTCAGA CAGTTATCAG TATGGAGAAG AAGTTACGTA CAAATGTTTT 420 GAAGGTTTTG GAATTGATGG GCCTGCAATT GCAAAATGCT TAGGAGAAAA AT 472 (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 385 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: GTATCTGTTC TTTGCCAAGA AAATTATCTA ATTCAGGAAG GGGAAGAAAT TACATGCAAA 60 GATGGAAGAT GGCAGTCAAT•ACCACTCTGT GTTGAAAAAA TTCCATGTTC ACAACCACCT 120 CAGATAGAAC ACGGAACCAT TAATTCATCC AGGTCTTCAC AAGAAATTTA TGCACATGGG 180 ACTAAATTGA GTTATACTTG TGAGGGTGGT TTCAGGATAT CTGAAGAAAA TGAAACAACA 240 TGCTACATGG GAAAATGGAG TTCTCCACCT CAGTGTGAAG GCCTTCCTTG TAAATCTCCA 300 CCTGAGATTT CTCATGGTGT TGTAGCTCAC ATGTCAGACA GTTATCAGTA TGGAGAAGAA 360 GTTACGTACA AATGTTTTGA AGGTT 385 (2) INFORMATION FOR SEQ ID NO: 35: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 385 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 35: GTATCTGTTC TTTGCCAAGA AAATTATCTA ATTCAGGAAG GAGAAGAAAT TACATGCAAA 60 GATGGAAGAT GGCAGTCAAT ACCACTCTGT GTTGAAAAAA TTCCATGTTC ACAACCACCT 120 CAGATAGAAC ACGGAACCAT TAATTCATCC AGGTCTTCAC AAGAAATTTA TGCACATGGG 180 ACTAAATTGA GTTATACTTG TGAGGGTGGT TTCAGGATAT CTGAAGAAAA TGAAACAACA 240 TGCTACATGG GAAAATGGAG TTCTCCACCT CAGTGTGAAG GCCTTCCTTG TAAATCTCCA 300 CCTGAGATTT CTCATGGTGT TGTAGCTCAC ATGTCAGACA GTTATCAGTA TGGAGAAGAA 360 GTTACGTACA AATGTTTTGA AGGTT 385 (2) INFORMATION FOR SEQ ID NO: 36: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 amino acids (B) TYPE: am o acid (C) STRANDEDNESS: (D) TOPOLOGY: linear ( x ) SEQUENCE DESCRI PTION : SEQ I D NO : 36 : Asn Cys Ser Met Ala Gin He Gin Leu Cys Pro Pro Pro Pro Gin He 1 5 10 15 Pro Asn Ser His Asn Met Thr Thr Thr Leu Asn Tyr Arg Asp Gly Glu 20 25 30 Lys Val Ser Val Leu Cys Gin Glu Asn Tyr Leu He Gin Glu Gly Glu 35 40 45 Glu He Thr Cys Lys Asp Gly Arg Trp Gin Ser He Pro Leu Cys Val 50 55 60 Glu Lys He Pro Cys Ser Gin Pro Pro Gin He Glu His Gly Thr He 65 70 75 80 Asn Ser Ser Arg Ser Ser Gin Glu Ser Tyr Ala His Gly Thr Lys Leu 85 90 95 Ser Tyr Thr Cys Glu Gly Gly Phe Arg He Ser Glu Glu Asn Glu Thr 100 105 110 Thr Cys Tyr Met Gly Lys Trp Ser Ser Pro Pro Gin Cys Glu Gly Leu 115 120 125 Pro Cys Lys Ser Pro Pro Glu He Ser His Gly Val Val Ala His Met 130 135 140 Ser Asp Ser Tyr Gin Tyr Gly Glu Glu Val Thr Tyr Lys Cys Phe Glu 145 150 155 160 Gly Phe Gly He Asp Gly Pro Ala He Ala Lys Cys Leu Gly Glu Lys 165 170 175 Trp (2) INFORMATION FOR SEQ ID NO:37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 123 ammo acids (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 37: Val Ser Val Leu Cys Gin Glu Asn Tyr Leu He Gin Glu Gly Glu Glu 1 5 10 15 He Thr Cys Lys Asp Gly Arg Trp Gin Ser He Pro Leu Cys Val Glu 20 25 30 Lys He Pro Cys Ser Gin Pro Pro Gin He Glu His Gly Thr He Asn 35 40 45 Ser Ser Arg Ser Ser Gin Glu He Tyr Ala His Gly Thr Lys Leu Ser 50 55 60 Tyr Thr Cys Glu Gly Gly Phe Arg He Ser Glu Glu Asn Glu Thr Thr 65 70 75 80 Cys Tyr Met Gly Lys Trp Ser Ser Pro Pro Gin Cys Glu Gly Leu Pro 85 90 95 Cys Lys Ser Pro Pro Glu He Ser His Gly Val Val Ala His Met Ser 100 105 110 Asp Ser Tyr Gin Tyr Gly Glu Glu Val Thr Tyr 115 120 (2) INFORMATION FOR SEQ ID NO: 38: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 123 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: linear (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38: Val Ser Val Leu Cys Gin Glu Asn Tyr Leu He Gin Glu Gly Glu Glu 1 5 10 15 He Thr Cys Lys Asp Gly Arg Trp Gin Ser He Pro Leu Cys Val Glu 20 25 30 Lys He Pro Cys Ser Gin Pro Pro Gin He Glu His Gly Thr He Asn 35 40 45 Ser Ser Arg Ser Ser Gin Glu He Tyr Ala His Gly Thr Lys Leu Ser 50 55 60 Tyr Thr Cys Glu Gly Gly Phe Arg He Ser Glu Glu Asn Glu Thr Thr 65 70 75 BO Cys Tyr Met Gly Lys Trp Ser Ser Pro Pro Gin Cys Glu Gly Leu Pro 85 90 95 Cys Lys Ser Pro Pro Glu He Ser His Gly Val Val Ala His Met Ser 100 105 110 Asp Ser Tyr Gin Tyr Gly Glu Glu Val Thr Tyr 115 120

Claims: CLAIMS 1. A method of screening for a cancer comprising the step of detecting the presence of a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding said antigen, said nucleic acid molecule characterized by the ability of said nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO: 18 and SEQ ID NO: 19, respectively). 2. A method according to claim 1 wherein the method comprises the step of detecting the presence of the antigen. 3. A method according to claim 1 wherein the method comprises the step of detecting the presence of a nucleic acid molecule encoding the antigen. 4. A method according to claim 1, 2 or 3 wherein the cancer is urogenital or renal cancer. 5. A method according to claim 4 wherein the urogenital cancer is bladder, cervical or prostate cancer. 6. A method according to claim 1, 2 or 3 wherein the antigen is further characterized by the ability to bind complement fragment C3b. 7. A method according to claim 1 , 2 or 3 wherein the antigen is further characterized by the ability to bind to heparin agarose. 8. A method according to claim 1, 2 or 3 wherein the antigen is further characterized by the presence of a polypeptide with a molecular weight of 138,000 which shifts to a molecular weight of 151 ,000 in the presence of a disulfide reducing agent, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 9. A method according to claim 1, 2 or 3 wherein the antigen is further characterized by at least two of: (1) the ability to bind complement fragment C3b, (2) the ability to bind to heparin agarose, and (3) the presence of a polypeptide with a molecular weight of 138,000 which shifts to a molecular weight of 151,000 in the presence of a disulfide reducing agent, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 10. A method of treating a tumor cell comprising the step of modulating a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding said antigen, said nucleic acid molecule characterized by the ability of said nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO: 18 and SEQ ID NO: 19, respectively). 1 1. A method according to claim 10 wherein the method comprises the step of modulating the antigen. 12. A method according to claim 10 or 11 wherein the tumor cell is a urogenital or renal tumor cell. 13. A method according to claim 12 wherein the urogenital tumor cell is a bladder, cervical or prostate tumor cell. 14. A method according to claim 10 or 11 wherein the antigen is further characterized by the ability to bind complement fragment C3b. 15. A method according to claim 10 or 11 wherein the antigen is further characterized by the ability to bind to heparin agarose. 16. A method according to claim 10 or 1 1 wherein the antigen is further characterized by the presence of a polypeptide with a molecular weight of 138,000 which shifts to a molecular weight of 151 ,000 in the presence of a disulfide reducing agent, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 17. A method according to claim 10 or 11 wherein the antigen is further characterized by at least two of: (1) the ability to bind complement fragment C3b, (2) the ability to bind to heparin agarose, and (3) the presence of a polypeptide with a molecular weight of 138,000 which shifts to a molecular weight of 151,000 in the presence of a disulfide reducing agent, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 18. An agent that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding said antigen, said nucleic acid molecule characterized by the ability of said nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO:19, respectively), for use as a medicament to treat a tumor cell. 19. A composition comprising an agent that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding said antigen, said nucleic acid molecule characterized by the ability of said nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO: 10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO: 19, respectively), in combination with a pharmaceutically acceptable carrier or diluent. 20. Use of an agent that modulates a tumor-associated human complement Factor H-related antigen or a nucleic acid molecule encoding said antigen, said nucleic acid molecule characterized by the ability of said nucleic acid molecule to hybridize under moderate stringency with the primer pair 42M/1040RT (SEQ ID NO:10 and SEQ ID NO: 17, respectively) or the primer pair 2910M/3610RT (SEQ ID NO:18 and SEQ ID NO:19, respectively), for the manufacture of a medicament for the treatment of a tumor cell.

Inventors: ENFIELD DAVID L, HASS G MICHAEL, KINDERS ROBERT J, ENFIELD, DAVID, L., HASS, G., MICHAEL, KINDERS, ROBERT, J.
Applicants: BARD DIAGNOSTIC SCIENCES INC, ENFIELD DAVID L, HASS G MICHAEL, KINDERS ROBERT J, BARD DIAGNOSTIC SCIENCES, INC., ENFIELD, DAVID, L., HASS, G., MICHAEL, KINDERS, ROBERT, J.
Application year: 1997
Main classification: C12Q1
IPC codes: C12Q1, G01N33