SEQUENTIAL TARGETING OF TUMOR SITES WITH OLIGONUCLEOTIDE CONJUGATES OF ANTIBODY AND COMPLEMENTARY OLIGONUCLEOTIDE CONJUGATES OF CHELATED RADIONUCLIDE

Abstract: The present invention is directed to a non-radioactive targeting immunoreagent that comprises oligonucleotides comprised of non-self-associating oligonucleotide sequences, and one or more linking groups and a radioactive targeting immunoreagent that comprises an oligonucleotide comprised of an oligonucleotide sequence that is complementary in sequence to and capable of hybridization with one or more fragments of a non-self-associating oligonucleotide sequence, one or more chelating agents, one or more linking groups and one or more radionuclides. The present invention is also directed to pharmaceutical compositions comprising one or more of the above-described immunoreagents and a pharmaceutically acceptable carrier. The present invention is further directed to methods for treating and imaging disease sites in patients.

Description: SEQUENTIAL TARGETING OF TUMOR SITES WITH OLIGONUCLEOTIDE CONJUGATES OF ANTIBODY AND COMPLEMENTARY OLIGONUCLEOTIDE CONJUGATES OF CHELATED RADIONUCLIDE Field of the Invention This invention relates to sequential targeting and delivery of radioactive immunoreagent compositions which find particular utility in therapeutic and diagnostic imaging methods. This invention also relates to novel methods for the attachment of oligonucleotides, complementary oligonucleotides, and chelates, to proteins, and to bifurcated tumor targeting and delivery vectors for the treatment and diagnostic imaging of tumors. Background of the Invention The various currently available radiolabeled immunoreactive proteins which are employed in diagnostic imaging and targeted therapeutic applications suffer from one or more of the following disadvantages: 1) toxicity; 2) destruction of the reagent due to rapid metabolism; 3) inadequate emission characteristics such as low signal to noise ratio; 4) inefficient cσvalent bonding with protein for conjugate preparation; 5) slow complexation with metals; 6) unstable metal complexation, e.g., with respect to temperature, time or pH; 7) inability to form conjugates until after metal complexation is accomplished; 8) inability to spectrophotometrically analyze the radionuclide complex reagent; 9) inability to form complexes without activation steps that degrade protein; UB 10) prolonged plasma half-lives of radioactive immunoconjugates result in prolonged exposure of normal tissue to damaging effects of radiation; 11) slow clearance of radionuclide from the body; and 12) increasing the number of sites of incorporation of radionuclide or chelated radionuclide results in a reduction in the capacity of the radiolabeled immunoreactive protein to bind to its target. It is an object of the present invention to overcome the aforementioned disadvantages of the currently available radiolabeled immunoreactive proteins. Summary of the Invention The present invention is directed to a non- radioactive targeting immunoreagent that comprises a tumor antigen recognizing moiety, one or more oligonucleotides comprised of non-self-associating oligonucleotide sequences, and one or more linking groups. The present invention is also directed to a radioactive targeting immunoreagent that comprises an oligonucleotide comprised of an oligonucleotide sequence that is complementary in sequence to and capable of hybridization with one or more fragments of a non-self- associating oligonucleotide sequence, one or more chelating agents, one or more linking groups and one or more radionuclides. The present invention is also diiected to pharmaceutical compositions comprising one or more of the above-described immunoreagents and a pharmaceutically acceptable carrier. The present invention is further directed to methods for treating and imaging disease sites such as tumor sites in a patient. Said methods comprise SUBSTITUTE SHEET administration to the patient of an effective amount of the above-described non-radioactive targeting immunoreagent followed at an effective time interval by an effective amount of the above-described radioactive targeting immunoreagent. The present invention provides many advantages compared to conventional targeting immunoreagents. For example, the non-radioactive targeting immunoreagent can accumulate at a tumor site in vivo while it is not accumulated at normal tissue sites. The in vivo residence half life of the non- radioactive targeting immunoreagent is long enough to permit its accumulation at a tumor site. The in vivo residence half life of the radioactive targeting reagent is shorter than the residence half life of the non-radioactive targeting immunoreagent. The portion of the radioactive targeting reagent that does not hybridize to tumor associated non- radioactive targeting reagent is rapidly cleared from the patient. With respect to the same degree of modification of a targeting immunoreagent by a directly labeled radionuclide or a chelate containing a radionuclide, an amplification of the number of radionuclides per site of modification per targeting immunoreagent can be obtained. A segment of the complementary sequenced oligonucleotides of the non-radioactive targeting immunoreagent and a segment of the radioactive targeting reagent can hybridize in vitro and in vivo. The complementary sequenced oligonucleotides of the non-radioactive targeting immunoreagent and the radioactive targeting reagent are stable to exonuclease activity. SUBSTITUTE SHE The complementary sequenced oligonucleotides of the non-radioactive targeting immunoreagent and the radioactive targeting reagent do not self hybridize. The non-radioactive targeting immunoreagent and the radioactive targeting reagent can comprise a wide variety of spacing, linking, and chelating groups, oligonucleotide sequences and radionuclides. The complementary sequenced oligonucleotides of the non-radioactive targeting immunoreagent can be comprised of oligonucleotide sequences which can be tandemly linked by spacing groups, wherein a segment of each oligonucleotide sequence can hybridize with a segment of the radioactive targeting reagent. The complementary sequenced oligonucleotides of the non-radioactive targeting immunoreagent can be linked to an antibody by means of either a 5'- or a 3'-substituent such as a 5'-amine or 3'-amine. Reagents are provided that have a specificity for tumors and a wide variety of compositions, size and molecular weight can be prepared in accordance with the present invention. A particular advantage of the present invention is that oligonucleotide sequence lengths and spacing groups can be selected such that on hybridization of two complementary radioactive targeting reagent oligonucleotide sequences to a single oligonucleotide- containing strand comprised of adjacent, tandemly linked oligonucleotide sequences of a non-radioactive targeting immunoreagent, the proximal end groups of the sequences of two such radioactive targeting moieties are orthogonal to each other because of their relative spacial configuration on the double stranded helix so formed. SUBSTITUTE SHEET Other advantageous features of this invention will become readily apparent upon reference to the following description of the preferred embodiments. Description of the Preferred Embodiments This invention describes various novel bioconjugates which possess utility in therapeutic and diagnostic imaging compositions and methods. This invention further describes novel methods of preparing bioconjugates by the attachment of various oligonucleotide sequences to chelating agents, preferably terpyridine containing chelating agents, and to immunoreagents such as proteins, antibodies, and receptors. In particular, this invention describes novel bioconjugates useful for sequential targeting and amplified delivery of novel radioactive immunoreagent compositions which find particular utility in therapeutic and diagnostic imaging compositions and methods. More particularly, this invention describes the preparation and use of targeting immunoreagents that comprise a tumor antigen recognizing moiety, one or more oligonucleotides comprised of non self-associating oligonucleotide sequences, and one or more linking groups. These targeting immunoreagents react with a radioactive sequential targeting reagent that comprises an oligonucleotide comprised of a oligonucleotide sequence that is complementary in sequence to and capable of hybridization with one or more fragments of the said non self-associating oligonucleotide sequences, one or more chelating agents, one or more linking groups, and having one or more radionuclides associated therewith. Most preferably, the nucleotides are deoxyribonucleotides. SUBSTITUTE SHEE In a preferred embodiment, the above-described targeting immunoreagents form a compound that comprises moieties represented by the structure 1: Structure 1 wherein: Z comprises the residue of an immunoreactive protein; L and 1^ are independently a chemical bond or a linking group; I comprises an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains one or more members of a family of homologous contiguous sequences, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units, and provided that contiguous sequences of six or more nucleotide units of said oligonucleotide do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure l; Qi is a spacing group; a is 0 or an integer from 1 to about 6; Ii comprises an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units, a contiguous sequence therein comprising a portion of I; E is an end capping group; and p is an integer from 1 to about 10. More preferably, a is an integer from 1 to about 6. SUBSTITUTE SHEE In another preferred embodiment, the above- described targeting reagent is comprised of moieties represented by the structure 2: Structure 2 wherein: cl comprises an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains therein one or more members of a family of homologous contiguous sequences, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units; where the nucleotide sequences of said members of said family of homologous contiguous sequences are complementary to the nucleotide sequences of members of the set of oligonucleotides in a coadministrable targeting immunoreagent and contiguous sequences of six or more nucleotide units of said complementary oligonucleotide do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure 2; Q CI is a spacing group; L ! , L 2 , and L 3 are independently a chemical bond or a linking group; W ιr W 2 , and W 3 are residues of chelating groups; SUBSTIT T HE j, M 2 , and M, are comprised of elements with oxidation states equal to or greater than +1, and at least one of M lf M 2 nd M 3 is a radionuclide; x, y, and z are independently zero or one provided that at least one of x, y, or z is one; and w and b are independently zero or an integer from l to about 4. The term "residue" as used herein in context with a chemical entity, which chemical entity is comprised of, for example, a chelating group, or a linking group, or a protein reactive group, or an immunoreactive group, or an immunoreactive protein, or an antibody, or an antibody fragment, or a cross-linking agent such as a heterobifunctional cross-linking agent, or an oligonucleotide, or a spacing group, or an end capping group, is defined as that portion of said chemical entity which exclusively remains when one or more chemical bonds of said chemical entity when considered as an independent chemical entity, are altered, modified, or replaced to comprise one or more covalent bonds to one or more other chemical entities. Thus, for example, in one aspect, a "residue of an oligonucleotide" in the context of, for example, I and Ii in Structure 1 or of cl in Structure 2 is comprised of an oligonucleotide modified at least for divalent attachment to the residue of another chemical entity, i.e., the residue of said oligonucleotide is comprised of at least a divalent oligonucleotidyl sequence. In another aspect, for example, "the residue of a chelating group" in the context of W α , W 2 or W 3 of Structure 2 is comprised of a chelating group which is at least monovalently modified through attachment to the residue of another chemical entity such as, for example, to the residue of a linking group. SUBSTITUTE SHEE In Structure 1 above, Z preferably is an antibody or antibody fragment which recognizes and is specific for a tumor associated antigen. In some embodiments, the above-described protein can contain an immunoreactive group covalently bonded thereto through a chemical bond or a linking group derived from the residue of a protein reactive group and the residue of a reactive group on the protein. As used herein, the term "immunoreactive protein" which can be abbreviated by "IRP" also includes an organic compound which is capable of covalently bonding to the protein and which is found in a living organism or is useful in the diagnosis, treatment or genetic engineering of cellular material or living organisms, and which has a capacity for interaction with another component which may be found in biological fluids or associated with cells to be treated such as tumor cells. Depending upon the intended use, the IRPs can be comprised of a wide variety of naturally occurring or synthetically prepared materials, including, but not limited to, in part or in their entirety, enzymes, amino acids, peptides, polypeptides, proteins, lipoproteins, lipids, phospholipids, glycoproteins, hormones, drugs (for example digoxin, phenytoin, phenobarbital, thyrozine, triiodothyronine, gentamicin, carbamazepine, and theophylline) , steroids, vitamins, polysaccharides, viruses, protozoa, fungi, parasites, rickettsia, molds, and components thereof, blood components, tissue and organ components, pharmaceuticals, haptens, lectins, toxins, nucleic acids (including oligonucleotides) , antibodies, antigenic materials (including proteins and carbohydrates) , avidin and derivatives thereof, biotin and derivatives thereof, and others known to one skilled in the art. Preferred IRPs are those which are comprised of the residue of a receptor molecule specific to the reεidue of a ligand of interest. Thus, a specific binding reaction involving the immunoreagent can be used for the targeting of a ligand of interest. Examples of such receptor-ligand combinations include, but are not limited to antibody-antigen, avidin-biotin, repressor (inducer) - promoter of operons and sugar-lectin complexes. Additionally, complementary nucleic acid sequences, i.e., a hybridized product of complementary strands, are also considered specific binding materials as the term is used herein. Useful IRPs are comprised of residues of substances which include (1) any substance which, when presented to an immunocompetent host., will result in the production of a specific antibody capable of binding with that substance, or (2) the antibody so produced, which participates in an antigen-antibody reaction. Thus, the IRP can be an antigenic material, an antibody, or an anti-antibody. Both monoclonal and polyclonal antibodies are useful. The antibodies can be whole molecules or various fragments thereof, as long as they contain at least one reactive site for reaction with the reactive groups on the precursors of the linking groups as described hereinbelow. As used herein, the term "antibody fragment" is used to describe any material derived from antibodies but which retains the ability to recognize and bind to an antigen. Such fragments may, for example, be produced by means of: (i) Enzymatic digestion (e.g., Fab or Fab' 2 fragments) ; (ii) Chemical means (e.g., the fragments produced by acid hydrolysis) ; or SUBSTITUTE SHEET (iii) Products created by molecular biology of or genetic engineering of the antibody genes (e.g. single chains or Fv fragments) . In certain embodiments, the IRP can be comprised of an enzyme which has a reactive group for attachment to the precursors of the linking groups as described hereinbelow. Representative enzymes include, but are not limited to, aspartate aminotransaminase, alanine aminotransaminase, lactate dehydrogenase, creatine phosphokinase, gamma glutamyl transferase, alkaline acid phosphatase, prostatic acid phosphatase, horseradish peroxidase and various esterases. If desired, the IRP can be modified or chemically altered to provide reactive groups for attaching to the residues of precursors of the linking groups or to oligonucleotides by techniques known to those skilled in the art. Such techniques include the use of linking moieties and chemical modification such as described in WO-A-89/02931 and WO-A-89/2932, which are directed to modification of oligonucleotides, and U.S. Patent No. 4,719,182. Two highly preferred uses for the compositions of this invention are for the diagnostic imaging of tumors and the radiological treatment of tumors. Preferred immunological groups therefore include antibodies or antibody fragments such as, for example. Fab, Fab'2, and single chain Fv fragments to tumor-associated antigens. Specific non limiting examples include B72.3 and related antibodies (described in U.S. Patent Nos. 4,522,918 and 4,612,282) which recognize colorectal tumors, 9.2.27 and related anti-melanoma antibodies, D612 and related antibodies which recognize colorectal tumors, UJ13A and related antibodies which recognize small cell lung carcinomas, NRLU-10 and related antibodies which recognize small cell lung carcinomas and colorectal tumors (Pan-Adenocarcinoma) , 7E11C5 and related antibodies which recognize prostate tumors, CC49 and related antibodies which recognize colorectal tumors, TNT and related antibodies which recognize necrotic tissue, PR1A3 and related antibodies, which recognize colon carcinoma, ING-1 and related antibodies, which are described in International Patent Publication WO-A- 90/02569, B174 and related antibodies which recognize squamous cell carcinomas, B43 and related antibodies which are reactive with certain lymphomas and leukemias, and anti-HLB and related monoclonal antibodies. An especially preferred antibody is ING-1. Referring to structure 1 again, L and 1^ are independently a chemical bond or the residue of a linking group. The phrase "residue of a linking group" as used herein refers to a moiety that remains, results, or is derived from the reaction of a protein reactive group with a reactive site on the protein. The phrase "protein reactive group" as used herein refers to any group which can react with functional groups typically found on proteins. However, it is specifically contemplated that such protein reactive groups can also react with functional groups typically found on nonprotein biomolecules. Thus the linking groups useful in the practice of this invention derive from those groups which can react with any biological molecule containing an immunoreactive group, whether or not the biological molecule is a protein, to form a linking group between the immunoreactive group and the oligonucleotide containing species as described below. Preferred linking groups are derived from protein reactive groups selected from but not limited to: (1) A group that will react directly with amine, alcohol, or sulfhydryl groups on the protein or biological molecule containing the immunoreactive group, SUBSTITUT SHEE for example, active halogen containing groups including, for example, chloromethylphenyl groups and chloroacetyl [C1CH 2 C(=0) -] groups, activated 2-(leaving group substituted)-ethylsulfonyl and ethylcarbonyl groups such as 2-chloroethylsulfonyl and 2-chloroethylcarbonyl; vinylsulfonyl; vinylcarbonyl; epoxy; isocyanato; isothiocyanato; aldehyde; aziridine; succinimidoxycarbonyl; activated acyl groups such as carboxylic acid halides; mixed anhydrides and the like; and other groups known to be useful in conventional photographic gelatin hardening agents; (2) A group that can react readily with modified proteins or biological molecules containing the immunoreactive group, i.e., proteins or biological molecules containing the immunoreactive group modified to contain reactive groups such as those mentioned in (1) above, for example, by oxidation of the protein to an aldehyde or a carboxylic acid, in which case the "linking group" can be derived from protein reactive groups selected from amino, alkyla ino, arylamino, hydrazino, alkylhydrazino, arylhydrazino, carbazido, semicarbazido, thiocarbazido, thiosemicarbazido, sulfhydryl, sulfhydrylalkyl, sulfhydrylaryl, hydroxy, carboxy, carboxyalkyl and carboxyaryl. The alkyl portions of said linking groups can contain from 1 to about 20 carbon atoms. The aryl portions of said linking groups can contain from about 6 to about 20 carbon atoms; (3) A group that can be linked to the protein or biological molecule containing the immunoreactive group, or to the modified protein as noted in (1) and (2) above by use of a crosslinking agent. The residues of certain useful crosslinking agents, such as, for example, homobifunctional and heterobifunctional gelatin hardeners, bisepoxides, and bisisocyanates can become a SUBSTITUTE SHEET part of, i.e., a linking group in, the protein- (oligonucleotide-containing species) conjugate during the crosslinking reaction. Other useful crosslinking agents, however, can facilitate the crosslinking, for example, as consumable catalysts, and are not present in the final conjugate. Examples of such crosslinking agents are carbodiimide and carbamoylonium crosslinking agents as disclosed in U.S. Patent No. 4,421,847 and the ethers of U.S. Patent No. 4,877,724. With these crosslinking agents, one of the reactants, such as the immunoreactive groμp, must have a carboxyl group and the other, such as the oligonucleotide containing species, must have a reactive amine, alcohol, or sulfhydryl group. In amide bond formation, the crosslinking agent first reacts selectively with the carboxyl group, then is split out during reaction of the thus "activated" carboxyl group with an amine to form an amide linkage between the protein and oligonucleotide containing species, thus covalently bonding the two moieties. An advantage of this approach is that crosslinking of like molecules, e.g., proteins with proteins or oligonucleotide containing species with themselves is avoided, whereas the reaction of, for example, homo- bifunctional crosslinking agents is nonselective and unwanted crosslinked molecules are obtained. Preferred useful linking groups are derived from various heterobifunctional cross-linking reagents such as those listed in the Pierce Chemical Company Immunotechnology Catalog - Protein Modification Section, (1991 and 1992) . Useful non-limiting examples of such reagents include: SUBSTITUTE SHEET Sulfo-SMCC Sulfosuccinimidyl 4-(N- maleimidomethyl)cyclohexane-1- carboxylate. Sulfo-SIAB Sulfosuccinimidyl (4- iodoacetyl)aminobenzoate. Sulfo-SMPB Sulfosuccinimidyl 4-(p- maleimidophenyl)butyrate. 2-IT 2-Iminothiolane. SATA N-Succinimidyl S-acetylthioacetate. In addition to the foregoing description, the linking groups, in whole or in part, can also be comprised of and derived from nucleotides and residues of nucleotides, both naturally occurring and modified. Particularly useful, non-limiting reagents for incorporation of modified nucleotide moieties containing reactive functional groups, such as amine and sulfhydryl groups, into an oligonucleotide sequence of this invention are commercially available from, for example, Clonetech Laboratories Inc. (Palo Alto California) and include Uni-Link AminoModifier (Catalog # 5190) , Biotin- ON phosphoramidite (Catalog # 5191) , N-MNT-C6- A inoModifier (Catalog # 5202) , AminoModifier II (Catalog # 5203), DMT-C6-3'Amine-ON (Catalog # 5222), C6-ThiolModifier (Catalog # 5211) , and the like. In one aspect, linking groups of this invention are derived from the reaction of a reactive functional group such as an amine or sulfhydryl group as are available in the above Clonetech reagents, one or more of which has been incorporated into an oligonucleotide sequence of this invention, with, for example, one or more of the previously described protein reactive groups such as heterobifunctional protein reactive groups, one or more of which has been incorporated into an immunoreagent of this invention. SUBSTITUTE SHEET Referring to Structure 1 again, I and Ii independently comprise an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains therein one or more members of a family of homologous contiguous sequences, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units, the individual homologs of said family of sequences, both individually or as a set of homologous sequences so comprised being hereinafter sometimes referred to as "the Sequence", and contiguous sequences of six or more nucleotide units of said oligonucleotide do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure 1. Members of the set of homologous contiguous sequences which comprise "the Sequence" can be found in both the sequence I and the sequence I ,* at least one such sequence is common to both I and Ii. The oligonucleotide sequence of I and Ii can be comprised of DNA, of RNA, or of purine and pyrimidine base modified DNA or RNA, or of backbone modified DNA or RNA such as methyl phosphonate or thiophosphonate or carbohydrate modified DNA or RNA analogs, whole or partially modified, or of combinations thereof as long as a complementary oligonucleotide sequence incorporated into the radioactive targeting moiety described below can hybridize to said oligonucleotide sequence to form a hybrid which exhibits a Tm (melting temperature) greater than about 37°C. Preferred oligonucleotides are DNA and RNA. Especially preferred is DNA comprised of A, T, G, and C nucleotide units. In a preferred embodiment, the oligonucleotide sequence I and Ii can be comprised of double stranded DNA or RNA. That is, the oligonucleotide sequence may be comprised of complementary DNA or RNA which forms a SUBSTITUTE SHEET double helix molecule. The complementary oligonucleotide sequence incorporated into the radioactive targeting moiety, composed of DNA or RNA, then hybridizes to one or the other of the strands of the double stranded DNA or RNA comprising I and Ii. In this way, the complementary oligonucleotide sequence incorporated into the radioactive targeting moiety interacts with the duplex DNA or RNA of I and Ii in such a way as to form triplex (triple helix) DNA, triplex RNA, or a triplex DNA-RNA hybrid. Preferred non-limiting examples of oligonucleotide sequences of which the "Sequence" can be comprised are shown below. The following sequences comprise a set of homologous oligonucleotide sequences which when considered individually or in any combination comprise a set herein defined as the "Sequence": (i) TTATGGACGGAG (SEQ ID NO:l); (ii) TTATGGACGGAGA (SEQ ID NO:2) ; (iii) TTATGGACGGAGAA (SEQ ID NO:3); (iv) TTATGGACGGAGAAG (SEQ ID NO:4); (v) TTATGGACGGAGAAGC (SEQ ID NO:5); (vi) TTATGGACGGAGAAGCT (SEQ ID NO:6); (vii) TTATGGACGGAGAAGCTA (SEQ ID NO:7); and (viii) TTATGGACGGAGAAGCTAA (SEQ ID NO:8). Of course, sequence (viii) contains sequence (vii) which contains sequence (vi) which contains sequence (v) , and so on. Thus, in structure 1, if I contains, for example sequence (iii) , and an Ii contains, for example, sequence (v) , then both I and I A contain at least sequence (iii) . Another Ii in structure 1 can contain (viii) , in which case it would also contain (i) through (vii) aε well as (viii) . In this case, all three sequences would contain at least (iii) [aε well as (i) and (ii)], and the two Ii'ε would contain at leaεt (v) [as well aε (i) through (iv) ] . In thiε regard, an SUBSTITUTE SHEET oligonucleotide that is comprised of a contiguous sequence of nucleotides, which sequence being complementary to at least sequence (i) , would hybridize to all sequences (i) through (viii) as would any member of a set of contiguous complementary sequences, the individual members of which are comprised of the sequence complementary to any of (i) through (viii) . Such a set of contiguous complementary sequences can comprise cl as will be described below. Another set of preferred homologous oligonucleotide sequences which can comprise the "Sequence" is: (ix) CGGAGAAGCTAA (SEQ ID NO:9); (x) ACGGAGAAGCTAA (SEQ ID NO:10); (xi) GACGGAGAAGCTAA (SEQ ID NO:11); ( ϋ) GGACGGAGAAGCTAA (SEQ ID NO:12); (Xiii) TGGACGGAGAAGCTAA (SEQ ID NO:13); (xiv) ATGGACGGAGAAGCTAA (SEQ ID NO:14); (xv) TATGGACGGAGAAGCTAA (SEQ ID NO:15); and (xvi) TTATGGACGGAGAAGCTAA (SEQ ID NO:8). An especially preferred sequence is: TTATGGACGGAGAAGCTAA (SEQ ID NO:8). Two or more of the oligonucleotide sequences of this invention can be tandemly linked by means of chemical bonds, by linking groups such as described above, or by spacing groups as described below. The sequential order of nucleotides in the oligonucleotide sequences of this invention can be from the 5' to the 3' end or from the 3' to the 5' end. Attachment of the oligonucleotide sequences of this invention via linking groups as described above to the immune reactive group as described above can be accomplished via 3' or via 5' sites or via derivatives attached to 3' or 5' εiteε of the oligonucleotide. As discussed elsewhere, the "Sequence" may also be composed of a double stranded DNA or RNA. That is, the SUBSTITUTE SHEET "Sequence" may consist of complementary oligonucleotides which noncovalently interact to form double stranded DNA or RNA. Attachment of this double stranded nucleic acid to the immune reactive group as described above can be accomplished via 3' or via 5' sites or via derivatives attached to 3' or 5' sites of the oligonucleotide. Referring to structure 1 again, Q τ is a spacing group which separates and links two or more oligonucleotide sequences of this invention. Q τ can be comprised of a linking group as defined above, alone, or in combination with a nucleotide or an oligonucleotide comprised of 2 to about 20 nucleotide units, the sequence of which is not self-associating or such that contiguous sequences of six or more nucleotide units comprised therein do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure 1. Q x can also be comprised of residues of an amino acid group, a peptide group, or a poly(alkylene oxide) group such as a poly(ethylene glycol) group. It is contemplated that each spacing group can be linked to from two to about six oligonucleotide sequences at least two of which containing the Sequence of this invention. Preferably, the spacing group is linked to two oligonucleotide sequenceε each of which contain the Sequence of thiε invention. Preferably, the spacing group is an oligonucleotide sequence. Non limiting examples of preferred spacing groups are oligonucleotides comprised of the following sequenceε: ACT; ACTC; ACTCT; CTC; TCTC; and SUBSTITUTE SHEET CTCTC . An eεpecially preferred εpacing group is an oligonucleotide: ACTCTC. Of course, the condition described above, i.e., that the oligonucleotide sequence of this invention does not comprise a self associating sequence, still applies when considering the selection of oligonucleotide spacing groups linked in combination with the oligonucleotide Sequence groups. In structure 1, a is from 0 to about 6, preferably an integer from 1 to about 6, more preferably one to about 4, and most preferably one or two. In structure 1, p is an integer from 1 to 10, preferably 1 to about 6, and more preferably 1 to 3. It is also contemplated that mixtures of immunoreactive proteins comprised of mixtures of Z modified as defined in structure 1 together with Z not so modified will also be useful in this invention. In this case, the bulk mixture properties of "p" of such mixtures would comprise fractional values from about zero to about 10. Preferably, the bulk p values would be from about 0.1 to about 10.0, more preferably from about 0.2 to about 5.0, and most preferably from about 0.4 to about 3. In structure 1, E is an end capping group. E is preferably a nucleotide group that is modified so as to reduce or prevent the action of exonuclease enzymatic activity on the oligonucleotide sequence. It can be a 3'- or 5'-phosphate linked ribose group containing one or more substituents such aε an alkyl group of 1 to about 10 carbon atomε, or an ether group such as alkyl or aryl or aralkyl or substituted aryl or aralkyl ether wherein the alkyl groups contain from 1 to about 10 carbon atoms and such alkyl or aryl groups may contain or be substituted by substituents containing oxygen, nitrogen or sulfur atoms, or a poly(alkylene oxidyl) SUBSTITUTE SHEET group, preferably at the 5'- or 3'- position, respectively, or elsewhere on the ribose group, which substituent will reduce or prevent the action of exonuclease enzymatic activity. A phosphate ester comprised of such entities is also useful, as well as a phoεphate eεter or modified ribose comprised of ele entε of a suitable linking group as defined above. E can also be comprised of Z or it can be attached to Q by elements of L as defined above to form a cyclic structure. E can also be comprised of compounds with a two carbon-one nitrogen atom internucleoside linkage. Preferably, E is comprised of a poly(alkylene glycol) phosphate diester. With respect to E, the poly(alkylene glycol) moiety can have from 2 to about 100 repeating units. Preferably, the poly(alkylene glycol) is a poly(ethylene glycol) . A currently preferred poly(alkylene glycol) phosphate diester is a tetra(ethylene glycol) phosphate diester, hereinafter sometimeε referred to as a "Teg" or "Teg unit". Such poly(alkylene glycol) phosphate diesters can be linked in tandem to each other to form a dimer phoεphate ester sequence, a trimer sequence, a tetramer sequence, and so forth. One or two such units is preferred. Such units can also be attached to residues of Q, L, I, or Z described above. A preferred end group E is comprised of a Teg unit linked by a phosphate ester bond to a nucleotide such as T. In the context of this invention, the term "modified nucleotide moiety" is intended to mean a chemical entity which is comprised of one or more chemical groups that are analogous to one or more portions of a naturally occurring nucleotide or of a residue of a naturally occurring nucleotide. A "modified nucleotide moiety" is comprised of that chemical entity which exclusively remains when one or SUBSTITUTE SHEET more chemical bonds, of which said naturally occurring nucleotide is otherwise comprised when considered as an independent chemical entity, is altered, modified, or replaced to comprise one or more covalent bonds to one or more other chemical entitieε, or iε compriεed of that chemical entity which excluεively remainε after removal or deletion of a portion, εuch aε, for example, a purine or pyrimidine base portion, a hydroxyl portion, a ribose portion, and the like or combinations thereof, of the naturally occurring nucleotide in one location simultaneouεly with said replacement of another portion of said nucleotide. Particularly useful, non-limiting examples of modified nucleotide moieties are comprised of reactive functional groups, such as amine and sulfhydryl groups. They can be commercially available such as, for example, those modified nucleotide moieties and precursors thereto which are available from Clonetech Laboratories Inc. (Palo Alto, California) . Said modified nucleotide moieties and precursors thereto include Uni-Link AminoModifier (Catalog #5190) , Biotin- ON phosphoramidite (Catalog #5191) , N-MNT-C6- AminoModifier (Catalog #5202), AminoModifier II (Catalog #5203) , DMT-C6-3'Amine-ON (Catalog #5222), C6- ThiolModifier (Catalog #5211) , and the like. One or more of εaid moieties can be incorporated into an oligonucleotide sequence comprising this invention. In one aspect, linking groups of this invention are derived from the reaction of a reactive functional group such as an amine or sulfhydryl group as are available in the above Clonetech reagents, one or more of which has been incorporated into an oligonucleotide sequence of this invention, with, for example, one or more of the previously described protein reactive groups such as heterobifunctional protein reactive groups, one or more SUBSTITUTE SHEET of which has been incorporated into an immunoreagent of this invention. A "modified nucleotide moiety" can comprise a nucleotide moiety that is modified so as to reduce or prevent the action of exonuclease enzymatic activity on the oligonucleotide sequence. It can be a 3'- or 5'- phosphate linked ribose or a 3'- or 5'-phosphate linked 2'-deoxyribose group containing one or more substituents such as an alkyl group of 1 to about 10 carbon atoms, or an ether group such as alkyl or aryl or aralkyl or substituted aryl or aralkyl ether wherein the alkyl groups contain from 1 to about 10 carbon atoms and such alkyl or aryl groups may contain or be substituted by subεtituentε containing oxygen, nitrogen or sulfur atoms, or a poly(alkylene oxidyl) group, preferably at the 5'- or 3'-ribose position, respectively, or elsewhere on the ribose group, which substituent will reduce or prevent the action of exonuclease enzymatic activity. A "modified nucleotide moiety" comprising a phosphate eεter comprised of said substituents is also useful, as well as a phosphate ester or modified ribose comprised of elementε of a εuitable linking group as defined above. A "modified nucleotide moiety" can also be comprised of Z or it can be attached to Q by elements of L as defined above to form a cyclic structure. A "modified nucleotide moiety" can also be compriεed of compounds with a two carbon-one nitrogen atom internucleoside linkage. Preferably, a "modified nucleotide moiety" iε compriεed of a poly(alkylene glycol) phosphate diester. With respect to said "modified nucleotide moiety", the poly(alkylene glycol) moiety can have from 2 to about 100 repeating units. Preferably, the poly(alkylene glycol) is a poly(ethylene glycol) . A currently preferred poly(alkylene glycol) phoεphate diester is a SUBSTITUTE SHEET tetra(ethylene glycol) phosphate diester, hereinafter εometi eε referred to aε a "Teg" or "Teg unit" . Such poly(alkylene glycol) phoεphate diesters can be linked in tandem to each other to form a dimer phosphate ester sequence, a trimer sequence, a tetramer sequence, and so forth. One or two such units is preferred. Such units can also be attached to residues of Q, L, I or Z described herein. A preferred "modified nucleotide moiety" iε compriεed of a Teg unit linked by a phosphate ester bond to a nucleotide such as T. Referring to the radioactive targeting immunoreagent described in Structure 2, preferred non- limiting examples of a set of oligonucleotides, cl, which are complementary to the members of the set of oligonucleotides comprising the "Sequence" of I in structure 1 include the DNA oligonucleotides: (xvii) TTAGCTTCTCCG (SEQ ID NO:16); (xviii) TTAGCTTCTCCGT (SEQ ID NO:17) ; (xix) TTAGCTTCTCCGTC (SEQ ID NO:18); (xx) TTAGCTTCTCCGTCC (SEQ ID NO:19); (xxi) TTAGCTTCTCCGTCCA (SEQ ID NO:20); (xxii) TTAGCTTCTCCGTCCAT (SEQ ID N0:21) ; (xxiii) TTAGCTTCTCCGTCCATA (SEQ ID N0:22); and (xxiv) TTAGCTTCTCCGTCCATAA (SEQ ID NO:23) Another set of preferred homologous oligonucleotide sequenceε which can compriεe a εet complementary to the "Sequence" of εtructure 1 iε: (xxv) CTCCGTCCATAA (SEQ ID NO:24) ; (xxvi) TCTCCGTCCATAA (SEQ ID NO:25) ; (xxvii) TTCTCCGTCCATAA (SEQ ID NO:26); (xxviii) CTTCTCCGTCCATAA (SEQ ID NO:27) ; (xxix) GCTTCTCCGTCCATAA (SEQ ID NO:28); (XXX) AGCTTCTCCGTCCATAA (SEQ ID NO:29) ; (xxxi) TAGCTTCTCCGTCCATAA (SEQ ID NO:30); (xxxii) TTAGCTTCTCCGTCCATAA (SEQ ID NO:23) SUBSTITUTE SHEET An especially preferred complementary sequence iε comprised of: TTAGCTTCTCCGTCCATAA (SEQ ID NO: 23) Of course, as is well known in the art, the complementarity of the above cl oligonucleotide sequences of structure 2 with respect to the previously listed I oligonucleotide sequences of structure 1 depends on the relative orientation of each, i.e., whether the sequenceε of I and of cl are constructed from 5' to 3' or 3' to 5', or vice versa, respectively. Preferably, the Tm of a hybridized complex formed between the respective I and cl sequences is greater than 37°C. In a preferred embodiment, the oligonucleotide sequence cl can be comprised of double stranded DNA or RNA. That iε, the oligonucleotide sequence may be comprised of complementary DNA or RNA which forms a double helix molecule. The complementary oligonucleotide sequence incorporated into the non- radioactive targeting immunoreagent, composed of DNA or RNA, then hybridizes to one or the other of the strands of the double stranded DNA or RNA comprising cl. In this way, the complementary oligonucleotide sequence incorporated into the non-radioactive targeting immunoreagent interacts with the duplex DNA or RNA of cl in such a way as to form triplex (triple helix) DNA, triplex RNA, or a triplex DNA-RNA hybrid. Q CI in structure 2 is a spacing group; Q cl can be selected from Q τ as described for structure 1. Preferably, Q CI is comprised of an oligonucleotide comprised of a contiguous sequence of from 2 to about 30 nucleotides, the sequence of which is not self- associating and such that a contiguouε εequence of six or more nucleotide units comprised therein does not hybridize with any other contiguous sequence of six or SUBSTITUTE SHEET more contiguous nucleotide units anywhere in structure 2. Q CI can also be comprised of an oligonucleotide, preferably a sequence such as (xxvii) to (xxxii) above which is complementary to the "Sequence" in structure l. Preferably, Q CI is comprised of one or two such sequences. l> l t L 2 , and L 3 in structure 2 are independently a chemical bond, preferably a phosphate ester bond, or a linking group which are defined as L and L-, in the above structure 1. L α , L 2 , and L 3 can also be independently comprised of components of Q cl . W : , W 2 , and W 3 in structure 2 are residueε of chelating groupε. The chelating groupε of thiε invention can compriεe the reεidue of one or more of a wide variety of chelating agentε that can have a radionuclide aεsociated therewith. As is well known, a chelating agent is a compound containing donor atoms that can combine by coordinate bonding with a metal atom to form a cyclic structure called a chelation complex or chelate. This class of compounds is described in the Kirk-Othmer Encyclopedia of Chemical Technology, Vol. 5, 339-368. The residueε of suitable chelating agentε can be independently εelected from polyphosphates, such as sodium tripolyphosphate and hexametaphosphoric acid; aminocarboxylic acids, such as ethylenediaminetetraacetic acid, N-(2- hydroxyethyl)ethylene-diaminetriacetic acid, nitrilotriacetic acid, N,N-di(2-hydroxyethyl)glycine, ethylenebis(hydroxyphenylglycine) and diethylenetriamine pentacetic acid; 1,3-diketoneε, εuch as acetylacetone, trifluoroacetylacetone, and thenoyltrifluoroacetone; hydroxycarboxylic acids, such as tartaric acid, citric acid, gluconic acid, and 5-sulfosalicylic acid; polyamines, such as ethylenediamine, diethylenetriamine, SUBSTITUTE SHEET triethylenetetramine, and triaminotriethylamine; aminoalcoholε, such as triethanolamine and N-(2- hydroxyethyl)ethylenediamine; aromatic heterocyclic baseε, εuch as 2,2'-dipyridyl, 2,2'-diimidazole, dipicoline amine and 1,10-phenanthroline; phenolε, such as salicylaldehyde, disulfopyrocatechol, and chromotropic acid; aminophenolε, such as 8- hydroxyquinoline and oximesulfonic acid; oximes, εuch as dimethylglyoxime and εalicylaldoxime; peptideε containing proximal chelating functionality such as polycysteine, polyhistidine, polyaεpartic acid, polyglutamic acid, or combinations of such amino acidε; Schiff bases, such as disalicylaldehyde 1,2- propylenediimine; tetrapyrroleε, εuch aε tetraphenylporphin and phthalocyanine; sulfur compounds, such aε toluenedithiol, meso-2,3- dimercaptoεuccinic acid, dimercaptopropanol, thioglycolic acid, potassium ethyl xanthate, sodium diethyldithiocarbamate, dithizone, diethyl dithiophoεphoric acid, and thiourea; synthetic macrocylic compounds, such as dibenzo[18]crown-6, (CH 3 ) 6 -[14]-4,ll-diene-N4, and (2.2.2-cryptate) ; and phosphonic acidε, εuch aε nitrilotrimethylene-phoεphonic acid, ethylenediaminetetra(methylenephoεphonic acid), and hydroxyethylidenediphoεphonic acid, or combinations of two or more of the above agents. Preferred residues of chelating agents contain polycarboxylic acid groups and include: ethylenediamine- N, N, N',N'-tetraacetic acid (EDTA) ; N,N,N',N'\N"- diethylene-triaminepentaacetic acid (DTPA) ; 1,4,7,10- tetraazacyclododecane-N,N' ,N",N"'-tetraacetic acid (DOTA) ; 1,4,7,10-tetraazacyclododecane-N,N' ,N"-triacetic acid (D03A) ; l-oxa-4,7,10-triazacyclododecane-N,N' ,N"- triacetic acid (OTTA) ; trans(l,2)- cyclohexanodiethylenetriamine pentaacetic acid (CDTPA) ; Preferred residues of chelating agents contain polycarboxylic acid groups and include the following: JMT DCDTPA PheMT MaαoPheMT MacroTMT In one aspect, other suitable residues of chelating agents are comprised of proteins modified for the chelation of metalε such as technetium and rhenium aε described in U.S. Patent No. 5,078,985, the disclosure of which is hereby incorporated by reference. In another aspect, suitable residues of chelating agents are derived from N 3 S and N 2 S 2 containing compounds, as for example, those disclosed in U.S. Patent Nos. 4,444,690; 4,670,545; 4,673,562; 4,897,255; 4,965,392; 4,980,147; 4,988,496; 5,021,556 and 5,075,099. Other suitable residueε of chelating agents are described in PCT/US91/08253 , the diεcloεure of which iε hereby incorporated by reference. In εtructure 2 above, if W- , W 2 , and W 3 are compriεed of the reεidue of multiple chelating agents, such agents can be linked . together by a linking group such as described above in structure 1. The residue of the chelating agents W lf W 2 , and W 3 in εtructure 2 are independently linked to the complementary oligonucleotide moiety cl or spacing group Q CI through a chemical bond or a linking group, i.e., L- , L 2 and L 3 in structure 2, above. Preferred linking groups include nitrogen atoms in groups εuch aε amino, imido, nitrilo and imino groups; alkylene, preferably containing from 1 to 18 carbon atoms such as methylene, ethylene, propylene, butylene and hexylene, such alkylene optionally being interrupted by 1 or more heteroatoms such as oxygen, nitrogen and sulfur or heteroatom-containing groups; carbonyl; sulfonyl ; sulfinyl; ether; thioether; eεter, i.e., carbonyloxy and oxycarbonyl; thioeεter, i.e., carbonylthio, thiocarbonyl, thiocarbonyloxy, and oxythiocarboxy; amide, i.e., iminocarbonyl and carbony1imino; thioamide, i.e., iminothiocarbonyl and thiocarbonylimino; thio; dithio; phoεphate; SUBSTITUTE SHEET phoεphonate; urelene; thiourelene; urethane, i.e., iminocarbonyloxy, and oxycarbony1imino; thiourethane, i.e., iminothiocarbonyloxy and oxythiocarbonylimino; an amino acid linkage, i.e., a group wherein n=l and X α , X 2 , X 3 independently are H, alkyl, containing from 1 to 18, preferably 1 to 6 carbon atoms, such as methyl, ethyl and propyl, such alkyl optionally being interrupted by 1 or more heteroatoms such as oxygen, nitrogen and sulfur, substituted or unsubstituted aryl, containing from 6 to 18, preferably 6 to 10 carbon atoms such as phenyl, hydroxyiodophenyl, hydroxyphenyl, fluorophenyl and naphthyl, aralkyl, preferably containing from 7 to 12 carbon atoms, such as benzyl, heterocyclyl, preferably containing from 5 to 7 nuclear carbon and one or more heteroatoms such as s, N, P or O, examples of preferred heterocyclyl groupε being pyridyl, quinolyl, imidazolyl and thienyl; heterocyclylalkyl, the heterocyclyl and alkyl portionε of which preferably are described above; or a peptide linkage, i.e., a group wherein n>l and each X independently is represented by a group as described for X-, X 2 , X 3 above. Two or more linking groups can be used, such as, for SUBSTITUTE SHEET example, alkyleneimino and iminoalkylene. It iε contemplated that other linking groups may be suitable for use herein, such as linking groups commonly used in protein heterobifunctional and homobifunctional conjugation and croεεlinking chemistry as described for L or L g in structure 1. Especially preferred linking groups include unsubstituted or substituted phosphate ester groups containing amino groups which when linked to the residue of a chelating agent via an isothiocyanate group on the chelating agent form a thiourea group. The linking groups can contain various substituents which do not interfere with the coupling reaction between chelate W-, W 2 , or W 3 and oligonucleotide cl or the spacing group. The linking groups can also contain substituents which can otherwise interfere with such reaction, but which during the coupling reaction, are prevented from so doing with suitable protecting groups commonly known in the art and which substituents are regenerated after the coupling reaction by suitable deprotection. The linking groups can also contain subεtituentε that are introduced after the coupling reaction. For example, the linking group can be substituted with substituents such as halogen, such as F, Cl, Br or I; an ester group; an amide group; alkyl, preferably containing from 1 to about 18, more preferably, 1 to 4 carbon atoms such as methyl, ethyl, propyl, i-propyl, butyl, and the like; substituted or unsubstituted aryl, preferably containing from 6 to about 20, more preferably 6 to 10 carbon atoms such as phenyl, naphthyl, hydroxyphenyl, iodophenyl, hydroxyiodophenyl, fluorophenyl and methoxyphenyl; substituted or unsubstituted aralkyl, preferably containing from 7 to about 12 carbon atoms, such aε benzyl and phenylethyl; alkoxy, the alkyl portion of SUBSTITUTE SHEET which preferably contains from 1 to about 18 carbon atomε aε described for alkyl above; alkoxyaralkyl, such as ethoxybenzyl; substituted or unsubεtituted heterocyclyl, preferably containing from 5 to 7 nuclear carbon and heteroatomε εuch aε S, N, P or 0, examples of preferred heterocyclyl groups being pyridyl, quinolyl, imidazolyl and thienyl; a carboxyl group; a carboxyalkyl group, the alkyl portion of which preferably contains from 1 to 8 carbon atoms; or the residue of a chelating group. In structure 2, M lf M 2 and M 3 are comprised of elements with oxidation stateε equal to or greater than +1, at leaεt one of which iε a radionuclide. Preferably Mj, M 2 and M 3 are compriεed of metal isotopes, preferably radioactive metal isotopeε, somethimes herein referred to aε metal radioiεotopeε, which radioisotopes are useful in therapeutic or in diagnostic imaging applications. Preferred metal radioisotopes are selected from, for example, Sc, Fe, Pb, Ga, Y, Bi, Mn, Cu, Cr, Zn, Ge, Mo, Tc, Ru, In, Sn, Re, Sr, Sm, Lu, Eu, Ru, Dy, Sb, W, Re, Po, Ta and Tl. Useful emisεionε from εuch radioiεotopes comprise spontaneous emissions comprised of alpha emissions, beta emisεionε, gamma emissions, X-ray emisεion, positron emisεionε, and comprise such emisεionε aε are induced by the processeε of electron capture and internal converεion. Said emiεεions can be purely of one kind such as pure alpha, pure beta, pure gamma and the like, or of combinations of nuclear emissions such as beta and gamma emissions and the like. Radioisotopeε with emiεεions compriεed, for example, of alpha radiation or of beta radiation are useful in therapeutic applications, especially in therapy of cancer. Useful isotopeε in therapeutic applications include, for example, alpha radiation SUBSTITUTE SHEET emitting iεotopes εuch as, for example, 207 Pb, 2U Pb, 08 Pb, 212 Pb, 212 Bi, 207 Ti, and 223 Ra; beta radiation emitting isotopes such as, for example 47 Sc, 66 Ga, 67 Cu, 77 As, 90 Y, 105 Rh, 109 Pd, nι Ag, 1 1 Sn, 127 Te, 143 Pr, 14 Pm, 153 Sm, 161 Tb, 166 Ho, 169 Er, 177 Lu, 188 Re, 186 Re, 191 0s and 199 Au; and iεotopeε which emit radiation as a result of the procesεes of electron capture and internal conversion such as, for example, 97 Ru, 177m Sn, 199 Sb, 128 Ba and 1 7 Hg. Radioisotopes especially preferred in therapeutic applications include 12 Pb, 21 Bi, 90 Y, 177 Lu, 186 Re, and 188 Re. Most preferred is 90 Y. Radioisotopes with emissionε comprised of, for example, gamma radiation or of positron radiation are useful in diagnostic imaging applications, especially in diagnostic imaging of cancer. Useful isotopes in diagnostic imaging applications include, for example, gamma radiation emitting isotopes such as 47 Sc, 51 Cr, 67 Cu, 67 Ga, 97 Ru, 99m Tc, ln, 117B Sn, 141 Ce, 167 Tm, 199 Au, 87 Y and 203 Pb; and positron radiation emitting isotopes εuch aε 44 Sc, 8 V, "Cu, 66 Ga, "Ge, 72 As, 86 Y and 89 Zr. Radioisotopeε eεpecially preferred in diagnoεtic imaging applicationε include "Cu, 99π Tc, ιn In and 87 Y. Moεt preferred are 99nι Tc and nι In. In another aεpect, other εuitable radionuclides can be incorporated, for example, by covalent bonding, into Qcl and include radioactive isotopeε of halogens such as radioactive isotopes of iodine, for example, 123 I, 124 I, 1 5 I and 131 I as well as radioactive isotopes of aεtatine such as 211 At. Methods of generating an image useful in the diagnostic imaging of, for example, cancer in a mammal comprise detecting emissions imagewise from radioisotopeε aε employed in the compoεitionε and methods of this invention. Said image generating methods comprise the uεe of, for example, a collimated SUBSTITUTE SHEET camera detector such as a gamma camera commonly employed in radioiitununoscintigraphy (RIS) , and the use of linked X-ray detectors commonly employed in positron emission tomography (PET) and in single photon emisεion tomography (SPET) . In structure 2, x, y, and z are independently zero or 1 provided that at least one of x, y, or z is one; and w and b are zero or an integer from 1 to about 4. Preferred compositions can be prepared as outlined in the schemes that follow. Scheme 1 Derivatization of antibody amine groups with heterobifunctional linking reagents SMCC, 2-IT, or SATA. In scheme 1, the protein (antibody, enzyme, receptor) is chemically modified for covalent coupling to a thiolated oligonucleotide or to a maleimido group containing oligonucleotide. Chemical modification iε effected uεing a bifunctional cross linking agent, preferably a heterobifunctional cross linking agent having both a group capable of reacting with protein functional groups (e.g. amine) and also having a further group capable of reacting with thiol groups. The latter is selected from haloacetyl, halo-acetamidyl, maleimido, and activated disulfide functions. Maleimido and thioalkyl groups are introduced to antibody by utilizing the heterobifunctional linkers, SUBSTITUTE SHEET sulfosuccinimido-4-(M-maleimidomethyl) -cyclohexane-1- carboxylate (Sulfo-SMCC) , 2-iminothiolane (2 IT) , or succinimidyl-S-acetylthioacetate(SATA) . The reaction of antibody with linking agent is for a time sufficient to introduce about 0.5-3 linking agent molecules per antibody molecule. The derivatized antibody is purified using a gel filtration column, and more preferably Sephadex G-25. Non-limiting examples of preferred protein conjugates prepared are listed below: ING-l-NH-CO-cyclohexane-CH 2 -Maleimide ING-1-NH-C(=NH)-(CH 2 )3-SH ING-l-NH-CO-CH 2 -SH Scheme 2 Derivatization of oligonucleotide "I-O^I" via amine groups with heterobifunctional linking reagents SATA and SMCC. o H δ-TEEG-l-Qrl -S —N S , TEG-l-Q,- l-3 ! — N V^SH & O O0go-3-- OGgα^'-SH SUBSTITUTE SHEE Scheme 3 Derivatization of oligonucleotide "I-Q--I" with amine groups and with sulfhydryl groups for uεe with heterobifunctional linking reagents. JTr Oligo-5'-SH 5ΗS-I-Q r l-NH 2 3' Scheme 4 Derivatization of oligonucleotide "δ'-TrS-I-Q j -I- 3'-NH 2 " with biotin for use with heterobifunctional linking reagents. TrS-δ'-OHgo-S'-NHa HS-5 # -l-Q r l-3'-Biotin HS-5'-Oligo-3*-Biotin SUBSTITUTE SHEET Scheme 5 Derivatization to produce "5'-HS-I-Q j -l-3'-biotin" with biotinylated phosphoramidite reagent for use with heterobifunctional linking reagents. O-DMT HS-5'-l-Q r I-3'-Biotin HS-5'-Oligo-3'-Biotin Oligonucleotides and modified oligonucleotides are synthesized according to standard methods known in the art. Derivatizations of oligonucleotides I-Q.-I were achieved utilizing the reaction of 5'-TEG-oligomer-NH 2 - 3' with SATA or SMCC affording oligomer-3'-SH or oligomer-3'-M (see Scheme 2). The following preferred oligonucleotides were used: 5'-Teg-TCTTATGGACGGAGAAGCTAAA TC CTTATGGACGGAGAAGCTAATCT -3'-amine-T, (SEQ ID No: 31; where the underlined portion denotes the spacer) ; and TTATGGACGGAGAAGCTAA (SEQ ID NO: 8) These oligomers are also derivatized to afford a bifunctionalized oligomer: 5'-HS-I-Q I -I-NH2-3' via introduction of bifunctional reagents at the 5'- and 3'-positions, as shown in Scheme 3. The introduction of biotin at the 3'-position is also achieved by the reaction of . δ'-HS-I-Q^I-NH,^' with biotin-i idocarboxylate aε εhown in Scheme 4. There are useful spacers with a protected amine, thiol, or carboxy on one end and a phosphoramidite at the other (εee Scheme 5) . The applications of these spacers are shown in reactions below. Scheme 6 Ab-M-5'-S-OIigo-3'-NH 2 I SUBSTITUTE SHEET Scheme 7 Assembly of Ab-SH + Oiigo-3'-M The modified maleimido antibody (2; Ab-M, Scheme 1) and the thiolated oligonucleotide {5 ' -nS-τ-Q τ -l-ϊlYi 2 -2 ' ) can be asεembled to yield the modified antibody- oligonucleotide conjugate (Ab-M-5'-S-succinimido-I-Q.-l- NH 2 -3') as shown in Scheme 6. Similarly, AB-S-M-3'-I- Q^I-TEG-5' iε prepared from AB-SH and δ'-TEG-I-Q ϊ -I- maleimide-3' aε εhown in Scheme 7. SUBSTITUTE SHEE Scheme 8 Complementary Oligonucleotide "5'-TMT-cI-TMT-3 N(iPr) 2 N F OC' (5') 2TMT-NCS 5'H 2 N-Cl-NH 2 3 * 5'T T-cI-T T 3' (3') wherein Ab = Antibody, protein; Oligo = oligonucleotide; cl = complementary oligonucleotide sequence; I -Qi-I =oligonucleotide; and TMT = terpyridine chelates. The reaction of 5'-H 2 N-cI-NH 2 -3' with the excess (2 mol) TMT-NCS affords the desired 5'-TMT-cI-TMT-3' as shown in Scheme 8. There are other useful linking agentε with a protected amine, thiol, or carboxy on one end and a phosphoramidite at the other. The applications of these agents are shown in the Examples. In a preferred embodiment, an effective dose of a radioactive targeting reagent as described above in a SUBSTITUTE SHEET pharmaceutically acceptable medium is prepared by exposing a composition of a complementary oligonucleotide sequence containing one or more chelating groups as described above to a composition containing a radioactive metal isotope such that the molar amount of said radionuclide metal isotope is less than the molar amount of said chelating group, said duration of exposure lasting an effective time to permit uptake of said metal isotope into said chelating agent. In a preferred embodiment, an effective dose of a non-radioactive targeting immunoreagent aε described above in a pharmaceutically acceptable medium is administered to a patient and said non-radioactive targeting immunoreagent is allowed to accumulate at the target site εuch aε at a tumor εite in εaid patient. Subsequently, an effective dose of a radioactive targeting reagent as described above in a pharmaceutically acceptable medium is administered to εaid patient, and said radioactive targeting reagent is allowed to accumulate at the target site, εaid target site being the non-radioactive targeting immunoreagent accumulated at said tumor site in said patient. The preεent invention includes one or more of the immunoreagents of this invention formulated into compositionε together with one or more non-toxic physiologically acceptable carriers, adjuvantε or vehicleε which are collectively referred to herein as carriers, for parenteral injection for oral administration in solid or liquid form, for rectal or topical administration, or the like. The compositions can be administered to humans and animals either orally, rectally, parenterally (intravenous, intramuscularly or subcutaneously) , intracisternally, intravaginally, intraperitoneally, SUBSTITUTE SHEE intravesically, locally (powders, ointments or drops) , or as a buccal or naεal spray. Compositions suitable for parenteral injection may comprise physiologically acceptable sterile aqueous or nonaqueous solutions, disperεions, suspensions or emulsions and sterile powders for reconstitution into εterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propyleneglycol, polyethyleneglycol, glycerol, and the like) , εuitable mixtureε thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersionε and by the use of surfactantε. Theεe compositions may also contain adjuvants such aε preεerving, wetting, e ulεifying, and dispensing agents. Prevention of the action of icroorganismε can be enεured by various antibacterial and antifungal agentε, for example, parabenε, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agentε, for example εugarε, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin. Solid dosage forms for oral administration include capsuleε, tabletε, pillε, powders and granules. In such solid doεage forms, the active compound is admixed with at least one inert customary excipient (or carrier) such as εodium citrate or dicalcium phoεphate or (a) fillers or extenderε, as for example, starcheε, lactoεe, εucroεe, glucose, mannitol and silicic acid, (b) binders, as for example, carboxymethylcelluloεe, SUBSTITUTE SHEET alignates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) hu ectants, as for example, glycerol, (d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicateε and sodium carbonate, (e) solution retarders, as for example paraffin, (f) absorption accelerators, as for example, quaternary ammonium compounds, (g) wetting agents, as for example, cetyl alcohol and glycerol monostearate, (h) adsorbents, as for example, kaolin and bentonite, and (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycolε, sodium lauryl sulfate or mixtures thereof. In the case of capsuleε, tablets and pills, the dosage forms may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethyleneglycols, and the like. Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shellε, εuch aε enteric coatingε and others well known in the art. They may contain opacifying agentε, and can alεo be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Exampleε of embedding compoεitionε which can be uεed are polymeric substances and waxes. The active compounds can also be in micro- encapsulated form, if appropriate, with one or more of the above-mentioned excipients. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suεpensions, εyrupε and elixirs. In addition to the SUBSTITUTE SHEET active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, εuch as water or other solvents ' , solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acid esterε of εorbitan or mixtureε of theεe εubεtanceε, and the like. Beεides such inert diluents, the composition can also include adjuvantε, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents. Suspensionε, in addition to the active compoundε, may contain suspending agents, as for example, ethoxylated isoεtearyl alcoholε, polyoxyethylene sorbitol and sorbitan esters, microcrystalline celluloεe, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substanceε, and the like. Compositions for rectal adminiεtrationε are preferably εuppositories which can be prepared by mixing the compounds of the present invention with εuitable non-irritating excipientε or carriers such as cocoa butter, polyethyleneglycol or a suppository wax, which are solid at ordinary temperatureε but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and releaεe the active component. Doεage formε for topical administration of a compound of this invention include ointments, powders, sprayε and inhalants. The active component is admixed under sterile conditions with a physiologically acceptable carrier and any preservativeε, bufferε or propellantε aε may be required. Ophthalmic SUBSTITUTE SHEET formulationε, eye ointments, powders and solutions are also contemplated as being within the scope of this invention. Actual dosage levelε of active ingredient in the compositions of the preεent invention may be varied so as to obtain an amount of active ingredient that is effective to obtain a deεired therapeutic response for a particular composition and method of administration. The selected dosage level therefore depends upon the desired therapeutic effect, on the route of administration, on the desired duration of treatment and other factorε. The total daily doεe of the compoundε of thiε invention adminiεtered to a hoεt in εingle or divided doεe may be in amountε, for example, of from about 100 picomol to about 5 micromolε per kilogram of body weight. Doεage unit compoεitionε may contain εuch amountε of εuch εubmultiples thereof as may be used to make up the daily dose. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the body weight, general health, sex, diet, time and route of administration, rates of absorption and excretion, combination with other drugs and the severity of the particular disease being treated. In another embodiment, the preεent invention is directed to a method of diagnosis comprising the adminiεtration of a diagnostic imaging effective amount of the compositionε of the present invention to a mammal in need of such diagnosiε. A method for diagnoεtic imaging for uεe in medical procedures in accordance with thiε invention compriεeε administering to the body of a test subject in need of a diagnostic image an effective diagnoεtic image producing amount of the above-deεcribed compoεitionε. SUBSTITUTE SHEET In this method, an effective diagnostic image producing amount of a non-radioactive targeting immunoreagent as described above in a pharmaceutically acceptable medium is administered to a patient and said non-radioactive targeting immunoreagent is allowed to accumulate at the target site such as at a tumor site in said patient. Subsequently, a diagnostic imaging effective dose of a radioactive targeting reagent as deεcribed above in a pharmaceutically acceptable medium is administered to said patient, and said radioactive targeting reagent is allowed to accumulate at the target site, said target site being the said non-radioactive targeting immunoreagent accumulated at said tumor site in said patient. The image pattern can then be visualized. In addition to human patients, the test subjects can include mammalian species such as rabbits, dogs, cats, monkeyε, εheep, pigε, horses, bovine animals and the like. After adminiεtration of the compositions of the present invention, the subject mammal is maintained for a time period εufficient for the adminiεtered compoεitionε to be distributed throughout the subject and enter the tissues of the mammal. A sufficient time period is generally from about 1 hour to about 2 weeks or more and, preferably from about 2 hours to about 1 week. The following examples further illustrate the invention and are not to be construed as limiting of the specification and claims in any way. Specific embodiments of the invention are illustrated in the following examples. SUBSTITUTE SHEET EXAMPLES Example 1 Oligonucleotide Design A targeting immunoreagent of this invention as described in Structure 1 comprised of an antibody Z, linking groupε L and L g , and an oligonucleotide comprised of a εequence I, a εpacing group Q-, a εecond sequence I lf and an end capping group E was designed as follows. A sample oligonucleotide with the following sequence (SEQ ID NO: 32) was analyzed for conformity to the criteria described previously for Structure 1 wherein groups L, I, Q τ r I i r and L g and E are as repreεented below: 5'TC]TTATGGACGGATCCGCTAA \TC{TTATGGACGGATCCGCTAA|TCT3 ' (SEQ ID NO 32) Analyεiε of thiε oligonucleotide sequence using "Oligo" computer softward (National Bioscienceε) revealed it to contain 5 regionε of self- complementarity. All εuch regionε were hairpin loops with differing degrees of overlap. The sum of the negative free energy changes (ΔG's) for these self- complementary regionε waε -28.3 kcal/mole which predicted a melting temperature of 86°C in 1.0 molar εalt for thiε oligonucleotide. The moεt εtable region of self-complementarity (ΔG = -15 kcal/mol) contained 8 base pairs most of which correspond to a BAM-1 restriction εite palindrome. In the preεence of theεe regionε of self-complementarity an oligonucleotide sequence complementary to thiε sequence cannot hybridize successfully to its full length. Modifications to thiε nucleotide sequence were then made in accordance with SUBSTITUTE SHEET the criteria outlined for εtructure 1 to replace εelected bases in regions of self-complementarity such that excessive base pairing was removed in iterative analysis. Additional nucleotides were also inserted into the spacer sequence to ensure that, in helical conformations, the terminal groups of two complementary sequences, cl, when hybridized to the two sequences I and I- were orthogonal to each other. In this manner, a new oligonucleotide (SEQ ID NO: 33) with the following εequence was designed, and then syntheεized and conjugated to an immunoreactive molecule (i.e., to an antibody): L I Q τ I τ Lc E 5X-TC TTATGGACGGAGAAGCTAA ACTCTC TTATGGACGGAGAAGCTAA TCTi.T-3 ' (SEQ ID NO: 33) This oligonucleotide (SEQ ID NO: 33) is comprised of two copies of the εame nucleotide sequence (i.e., I = Ii = TTATGGACGGAGAAGCTAA; SEQ ID NO: 8) linked through 6 nucleotide spacer (Q x = ACTCTC) . The groups L and I^E at the 5' or the 3' end are comprised of groupε X and Y which can be compriεed of either an amine-containing group (such as, for example, from those available from Clontech Industrieε, which amine-containing groupε being εometimeε hereinafter cryptically referred to aε "NH 2 " or as "amine") or one or more TEG groups as described above. Thus, in one embodiment, this oligonucleotide, cryptically referred to as S'-Teg-I-Qj-I-S'-amine, is: 5'Teg-TCTTATGGACGGAGAAGCTAAACTCTCTTATGGACGGAGAAGCTAATCT-3'-NH--T (SEQ ID NO: 31) 5'-Teg-I-Q- f -I-3'amine. SUBSTITUTE SHEET In another embodiment, thiε oligonucleotide, cryptically referred to aε δ'-amine-I-Q j -I-S'Teg, is: 5'H-N-TCTTATGGACGGAGAAGCTAAACTCTCTTATGGACGGAGAAGCTAATCT-3'-Teg-T (SEQ ID NO: 34) S'-amine-I-Q j -I-S'-Teg. Example 2 (2a) Svntheεis of I-O τ -I; 5'-Teα-I-0*,-I-3'-NH. (SEQ ID NO: 31) The oligonucleotide was prepared on an Applied Biosyste s oligonucleotide synthesizer by the trityl-on protocol as directed by the equipment manufacturer using 2-deoxynucleotide phosphoramidite reagent precursorε (5'-dimethoxytrityl cytidine-3'-O-phoεphoramidite, 5'- dimethoxytrityl adenoεine-3'-O-phoεphoramidite, 5'- dimethoxytrityl guanosine-3'-O-phosphoramidite, and 5'- dimethoxytrityl thymidine-3'-O-phosphoramidite from Applied Biosyεtemε) . Clonetech's Uni-link Amino Modifier was used aε the precurεor to the 3'-amine group. The TEG group, a (tetra(ethylene glycol)) phoεphate dieεter linked by a phoεphate eεter bond to 5'-dimethoxytrityl thymidine-3'-O-phoεphoramidite aε disclosed in WO/92/02534 was used at the 5'-end of the strand. Following synthesis of the whole oligonucleotide, the base protecting groups and solid support were removed with ammonium hydroxide and the resulting 5' protecting group was removed with 3% trichloroacetic acid. The oligonucleotide was desalted and further purified by elution down an OPEC Cartridge (Clonetech) with deionized water. Electrophoresis on a 12% polyacrylamide gel was used to further purify the oligonucleotides into upper and lower bands. The DNA band waε visualized by U.V. shadowing. It was cut out, minced, and extracted SUBSTITUTE SHE with a buffer comprised of lOmM Tris HC1 and ImM EDTA at pH 7.5 at 4"C for 24 hours. The gel pieces were then removed by centrifugation, and the DNA was purified through a spun column of Sephadex G-25. The concentration of oligonucleotide was estimated using absorbance at 260 nm. (2b) Synthesis of 5'-H ; N-I-O τ -I-3'-Tea (SEQ ID NO: 34) This oligonucleotide was prepared on an Applied Biosystemε oligonucleotide synthesizer by the trityl-off protocol as directed by the equipment manufacturer using 2-deoxynucleotide phosphoramidite reagent precursors (5'-dimethoxytrityl cytidine-3'-O-phosphoramidite, 5'- dimethoxytrityl adenosine-3'-O-phoεphoramidite, 5'- dimethoxytrityl guanoεine-3'-O-phosphoramidite, and 5'- dimethoxytrityl thymidine-3'-O-phosphoramidite from Applied Biosystems) . Clonetech'ε 6 carbon monomethoxytrityl AminoModifier (N-MMT-C6-AminoModifier) waε used as the precursor of the 5'-amine group. The TEG group was used as the terminal group at the 3' end. The base protecting group and εolid support were removed with ammonium hydroxide and the oligonucleotide waε further purified by polyacrylamide gel electrophoreεis. (2c) Svnthesiε of 5'-Trityl-S-I-O τ -3'-NH. The oligonucleotide εequence of Example 2a minus the Teg group was prepared on an Applied Biosystems oligonucleotide syntheεizer by the trityl-on protocol as directed by the equipment manufacturer uεing 2- deoxynucleotide phoεphoramidite reagent precurεorε (5'- dimethoxytrityl cytidine-3'-O-phoεphoramidite, 5'- dimethoxytrityl adenoεine-3'-O-phosphoramidite, 5'- dimethoxytrityl guanosine-3'-O-phosphoramidite, and 5'- SUBSTITUTE SHEET dimethoxytrityl thymidine-3'-O-phoεphoramidite from Applied Bioεystems) . Clonetech's Uni-link Amino Modifier waε uεed aε the precurεor to the 3'-amine group. Clonetech'ε C6-ThioModifier waε uεed as the precursor to the 5'-thiol group (which in herein referred to cryptically as 5-S) added to the sequence in place of Teg in Example 2a. Following synthesiε of the whole oligonucleotide, the baεe protecting groupε and solid support were removed with ammonium hydroxide. The oligonucleotide was desalted and further purified by elution down an OPEC Cartridge (Clonetech) with deionized water. The concentration of oligonucleotide was estimated using absorbance at 260 nm. (2d) Preparation of fluoreεcent S'Teq-I-O j -I-S '-NH- Cv5.18 A sample (30 nmoles) of δ'-Teg-I-Q- j -I-S'-NH 2 , prepared according to Example 2a, waε evaporated to dryneεε and rediεsolved in 500 μL 0.1 M bicarbonate buffer at pH9 by vortexing. This sample was then added to a vial containing the dried succinimidyl ester of the dye Cy5.18 (Biological Detection Systemε; Pittsburg PA) . After thorough mixing the reaction was allowed to proceed at room temperature for one hour with frequent mixing. The product, δ'-Teg-I-Q- r -I-S'-NH-CyS.18, was purified by elution from a Sepahadex G-25 column with distilled water. Example 3 (3a) Svntheεiε of cl diamine. 5'-NH,-cI-3'-NH : An oligonucleotide, cl, containing the nucleotide sequence 5'X-TTAGCTTCTCCGTCCATAAYT-3 ' (SEQ ID SUBSTITUTE SHEET NO: 35) complementary to I was prepared on an Applied Biosystems oligonucleotide synthesizer as outlined in Example 2a. The 3' (Y) and 5' (X) amine-containing groups were incorporated as directed by the equipment manufacturer using Uni-link Amino Modifier (Clonetech) for the precursor to the 3'-amine group, and Clonetech's 6 carbon monomethoxytrityl AminoModifier (N-MMT-C6- AminoModifier: Catalog # 5202) as precursor to the 5'- amine group. After final deblocking and removal from the solid support, the protecting groups were removed with ammonium hydroxide, the amine-functionalized oligonucleotide waε purified by elution down an OPEC Cartridge (Clonetech) with deionized water. The oligonucleotide was further purified by polyacrylamide gel electrophoresiε or reverεe-phase HPLC. The concentration of oligonucleotide was estimated uεing abεorbance at 260 nm. (3b) Svntheεiε of I diamine. 5'-NH,-I-3'-NH An oligonucleotide, I, containing the nucleotide sequence 5'X-TTATGGACGGAGAAGCTAAYT-3 ' (SEQ ID NO: 36) was prepared on an Applied Biosystems oligonucleotide syntheεizer as outlined in Example 3a. The 3' (Y) and 5' (X) amine-containing groups were incorporated as directed by the equipment manufacturer using Uni-link Amino Modifier (Clonetech) for the precursor to the 3'- amine group, and Clonetech's 6 carbon monomethoxytrityl AminoModifier (N-MMT-C6-AminoModifier: Catalog # 5202) as precursor to the 5'-amine group. After final deblocking and cleavage from the solid support, the protecting groups were removed with ammonium hydroxide, the amine-functionalized oligonucleotide was purified by elution down an OPEC Cartridge (Clonetech) with deionized water. The oligonucleotide was further SUBSTITUTE SHEET purified by polyacrylamide gel electrophoresis or reverse-phase HPLC. The concentration of oligonucleotide was estimated using absorbance at 260 nm. (3c) Synthesis of cl An oligonucleotide, cl, containing the nucleotide sequence 5'-TTAGCTTCTCCGTCCATAA-3 ' (SEQ ID NO: 23) complementary to (I) was prepared on an Applied Biosyεtemε oligonucleotide synthesizer as outlined in Example 2a. After final deblocking and cleavage from the solid support, the protecting groups were removed with ammonium hydroxide, and the oligonucleotide was purified by elution down an OPEC Cartridge (Clonetech) with deionized water. The oligonucleotide was further purified by polyacrylamide gel electrophoresis. The concentration of oligonucleotide was eεti ated uεing absorbance at 260 nm. (3d) Svnthesiε of I An oligonucleotide, I, containing the nucleotide εequence 5'-TTATGGACGGAGAAGCTAA-3' (SEQ ID NO: 8) was prepared on an Applied Bioεyεtemε oligonucleotide synthesizer as outlined in Example 3c. After final deblocking and cleavage from the solid support, the protecting groups were removed with ammonium hydroxide, and the oligonucleotide was purified by elution down an OPEC Cartridge (Clonetech) with deionized water. The oligonucleotide was further purified by polyacrylamide gel electrophoreεis. The concentration of oligonucleotide was estimated using absorbance at 260 nm. SUBSTITUTE SHEET (3e) Preparation of fluorescent cl and I A εample (30 nmoles) of cl diamine, prepared according to Example 3a, waε evaporated to dryneεε and rediεsolved in 500 μL 0.1 M bicarbonate buffer at pH9 by vortexing. This sample was then added to a vial containing the dried succini idyl ester of the dye Cy5.18 (Biological Detection Systems; Pittsburg PA). After thorough vortexing the reaction was allowed to proceed at room temperature for one hour with frequent mixing. CY5.18 labeled cl was purified by elution from a Sepahadex G-25 column with distilled water. A sample of Cy5.18 labeled I diamine (Example 3b) waε prepared in the εame manner. Example 4 Preparation of 3 P radiolabeled oligonucleotides Solutions (50 pmoles DNA in deionized water) of oligonucleotide (cl) from Example 3c or oligonucleotide (I) from Example 3d, were diluted into a buffer containing 0.5m Tris (pH 7.6), 0.1m MgCl 2# 50 mM dithiothreitol, ImM εpermine and ImM EDTA at room temperature. A sample (20 units) of T4 polynucleotide kinaεe waε added to each sample which was then incubated at 37'C. Radiolabeling was initiated by the addition of 50 pmoles of [γ - 32 P]ATP (3 Ci/μmole) to give a final total volume of 50 μL of each sample. After 60 minutes the reaction was halted by the addition of 2 μL of 0.5M EDTA. The radiolabeled DNA was extracted once with phenol/chloroform, precipitated with ice-cold ethanol and redisεolved in 50 mMTriε/EDTA buffer (pH 7.9) . SUBSTITUTE SHEET Finally the 32 P-cI or 32 P-I were passed down a Sephadex G-50 column to remove any unincorporated [γ - 3 P]ATP. Example 5 Hybridization of 32 P radiolabeled cl or I to I-O j -I A 5 pmole εample 32 P-labeled cl solution (from Example 4) was mixed with increasing amounts (0.375 to 12 pmoleε) of I-Q- t -I (from Example 2a) in PBS at 37'C for one hour. A 5μL aliquot from each of these hybridizations was removed, mixed with SDS buffer, and run on a 12% PAGE gel. Autoradiographs of the gels revealed that each I-Q τ -I waε capable of binding to 2 moleculeε of cl. Identical experiments using non- complementary, 3 P-labeled I (Example 4) failed to show hybridization to I-Q--I. Example 6 (6a) Annealing of cl to I-O j -I A sample of I-Qj-I (2 μmoles) , prepared according to Example 2a, was mixed with increasing amounts of cl (100:5 to 100:200, I-Q^IrcI), prepared by the method of Example 3c, in 6SSC buffer at room temperature. The mixtures were loaded into a sample cuvette and analysed by UV light (260 nm) in a Cary 13 instrument while the cuvette temperature was ramped up from 30"C to 90"C and then back down to 30°C. Analysis of the data revealed the presence of binary complexes (c^I-Q j -I and/or I-Q τ - I:cl) at concentrations of cl below equimolar with respect to I-Q Σ -I and with a melting tranεition temperature (TJ of around 48"C. At higher SUBSTITUTE SHEET concentrations of cl, a ternary complex (cl:I-Q j -I:cl) with a T m of 72.5°C could be seen. (6b) Hybridization of TMT-cI-TMT to I-O τ -I Similar experiments using cI-TMT (see Example 8a below) demonstrated the annealing of TMT-cI-TMT to I- Q z -I with a ternary complex T m of 74°C. Example 7 Preparation of 3S S-labeled TMT-NCS A suεpenεion of about 40 μmoleε of TMT amine (PCT US91/08253) in 650 μL methanol iε εtirred at room temperature and deionized water iε added dropwise (about 70 μL) until a clear pale yellow solution develops. The solution is cooled in an ice bath to 10"C and about 60 μmoles 35 S-thiophoεgene is added dropwise over about 3 minutes. A precipate of TMT isothiocyanate forms and the solution is stirred continuously for a f rther 2.5 hours. The solution and precipitate are concentrated to near drynesε on a rotovap under reduced preεsure ( - 15mm Hg) at room temperature. The near-dry solid iε diluted further with about 500 - 750 μL methanol and is εtirred until the solids appear homogeneous. The solids of TMT isothiocyanate are then collected by filtration and rinsed repeatedly with methanol. The product iε dried overnight in a vaccum chamber. SUBSTITUTE SHEET Example 8 (8a) Preparation 5 ' -H.N-cI-3'-NH-, conjugated to a chelating agent, TMT. to form TMT-cI-TMT. To 300 nmoles oligonucleotide 5'-H 2 N-cI-3'-NH 2 solution of Example 3a in 500 ul 1.0M carbonate/bicarbonate buffer at pH 9.0 was added 12 mg of TMT isothiocyanate (PCT US91/08253) . The reaction mixture was vortex mixed and kept at 37°C for 2 hours and at room temperature for overnight. The resulting reaction mixture was quenched with ethanolamine (15 uM) and the product waε purified by Sephadex G-25 column chromatography, eluting with deionized water. The number of TMTε per molecule of cl diamine was quantified by an aεssay using the time resolved fluorescence of chelated Europium metal. (8b) General procedure for labelling of Ing-l-TMT and TMT-cI-TMT conjugates with fluorescent metals. Binding of lanthanides εuch aε europium (3+) to chelating agentε that contain an aromatic moiety held cloεe to the co-ordination sphere can lead to "sensitized" fluorescence wherein light is absorbed through the aromatic εyεtem and the energy iε tranεferred to the metal. The metal then produces emiεεions characterized by a very large Stokeε εhift and fluorescence lifetimes of up to several seconds. The fluorescence at 615 nm is measured at a time-delay of 400 microseconds after an excitation pulεe at 340 nm. Thiε time delay is critical for high sensitivity measurements since short lived background fluorescence iε efficiently eliminated, and up to a 1,500 fold SUBSTITUTE SHEET enhancement in senεitivity over normal Eu-fluorescence is achievable. In this method, a known amount of Ing-1-TMT or TMT- cI-TMT conjugate is titrated with increasing amounts of added EuCl 3 in an aqueous buffer. Thus, one microliter of a solution containing 1-30 picomoles (0.15 μg - 5μg) of the conjugate is added, in duplicate, to wells in a Coεtar EIA/RIA 96-well plate containing a precalculated amount of Triε.HCl buffer (pH 7.4). The buffer volume is derived by subtracting from 99 the volume in microliters of aqueouε EuCl 3 (typically 10 ~4 M to 10 "6 M in Triε.HCl buffer). The total volume in each well is thereby fixed at 100 microliters. Aqueous EuCl 3 is then added to the buffered solution of the conjugate. The plate is then covered and shaken at low speed for one hour. The time reεolved fluoreεcence iε then meaεured using a Delfia 1232 time-resolved fluorimeter (Wallac Inc.) and the data are analyzed. It is found that each conjugated TMT molecule chelates one Europium ion and that the TMT-cI-TMT conjugate bindε two Europium ions per molecule of conjugate. (8c) Preparation of 5'-NH-,-cI-3'-NH-, Conjugated to 35 S-labeled TMT-NCS 35 S-labeled TMT-NCS( prepared as in Example 7) iε εubεtituted for the 12 mg of TMT iεothiocyanate (PCT US91/08253) in the method of Example 8a and the reaction carried out aε deεcribed above. The number of TMTε per molecule of cl is quantified by counting the TMT-cI-TMT product in a liquid scintillation counter optimized to detect 35 S. From a knowledge of the concentration of cl and the specific activity of 35 S-thiophosgene, the number of TMT moleculeε per cl diamine may be calculated. SUBSTITUTE SHEET Example 9 (9a) Preparation of a Yttrium ( 90 Y) radiolabeled- oliσonu ' cleotide. 90 Y-TMT-cI-TMT- 90 Y. A εolution of the oligonucleotide TMT conjugate (TMT-cI-TMT) from Example 8a in deionized water at room temperature waε treated with a εolution of 90 YC1 3 (>500 Ci/mg: Amersham) in 0.5 M sodium acetate buffer at pH 6.0 to a specific activity of 0.1 Ci/pmole for one hour at room temperature. The labeling efficiency was determined by removing 1.0 μL of the sample and spotting it on to a Gelman ITLC-SG strip. The strip was developed in a glasε beaker containing 0.1 M sodium citrate, pH 6.0 for a few minutes until the solvent front reached three-quaters of the way to the top of the paper. The strip waε then inεerted into a Syεtem 200 Imaging Scanner (Bioεcan) which had been optimized for 90 Y and waε controlled by a Compaq 386/20e computer. In thiε εyεtem unbound 90 Y migrateε at the solvent front . The TMT-cI-TMT chelated in excess of 98 % of the added radioactivity. (9b) Administration of 0 Y-TMT-cI-TMT- 9 °Y to Nude Mice TMT-cI-TMT, labeled with 90 Y to a specific activity of 28 μCi/28 μg, was injected into a 25g nude mouse bearing a subcutaneous tumor in its right flank. At time intervalε after injection, blood εamples were taken from the tail and counted for 90 Y radioactivity in a liquid εcintillation counter. The results revealed that more than 99% of the injected dose of 90 Y was removed from the blood stream in the first 30 minutes following SUBSTITUTE SHEET injection. After 2 hours following injection the radioactivity in the blood leveled off and a minute fraction ( < 0.01% of injected dose ) continued to circulate during the next 22 hours. These data confirm that, in the absence of a hybridizing site recognizing the cl sequence, radiolabeled cI-TMT iε loεt quickly from the circulation. Example 10 (10a) Preparation of Antibodv-Maleimide with Sulfo- SMCC fING-1-Maleimide) A Sulfo-SMCC solution (108 nmoles) in PBS waε added to a sample of a chimeric antibody (ING-1; 18 nmoles) εolution in phosphate buffer (pH7) . The resulting mixture waε allowed to εtand for 30 minuteε with occasional mixing at room temperature. The reaction waε stopped with 60 nmoles basic tris buffer. The reaction mixture was diluted with phoεphate buffered saline, added to a prewashed PD-10 column, and eluted with PBS to afford ING-1-maleimide. Thiε material waε stored on ice until use. (10b) Preparation of mercaptoalkyl-Antibodv fING-1-SH) A sample of a chimeric antibody (ING-1; 5 nmoles) εolution in 0.1 M carbonate buffer (pH 8.8) was mixed with 200 nmoles of an aqueous solution of 2- iminothiolane. The resulting mixture was allowed to stand for 30 min. with occasional mixing at room temperature. The reaction mixtures were diluted with phosphate buffed saline, added to a prewashed PD-10 column (Pharmacia) , and eluted with PBS to afford SUBSTITUTE SHEET mercaptoalkyl-ING-1. Thiε .material was stored on ice until use. Example 11 (11a) Preparation of a mercaptoalkyl-I-O τ -I using 2- iminothiolane, 5'-Teg-I-0 T -I-3'-SH A sample of a δ'-Teg-I-Q j -I-S'-NH 2 (30 nmoles) solution in water was mixed with 1 M carbonate buffer (pH 9) to give a final buffer concentration of 890 mM. Into the buffered DNA was added 12 μmoles of an aqueous solution of 2-iminothiolane and these reactants were vortex mixed and kept at 37°C for 30 minutes. The reaction mixture was quenched by the addition of 12 μmoles of ethanolamine, diluted with phosphate buffed saline, added to a prewashed NAP-25 column (Pharmacia) , and eluted with PBS to afford 5'Teg-I-Q j -I- 3'NHC(NH 2 + )CH 2 CH 2 CH 2 SH. For use in conjugation to maleimide-derivatized antibody, the product was eluted off the column directly into the antibody solution. Otherwise, mercaptoalkyl-I-Q j -I was stored on ice until use. (lib) Preparation of a mercaptoalkyl-I-O j -I using 2- immothiolane. 5'-HS-I-0 T -I-3'-Teg A sample of 5'-H 2 N-I-Q x -I-3'-Teg (30 nmoles) was treated as in Example 11a to afford 5'- HSCH 2 CH,CH 2 (NH 2 + )CHN-I-Q^I-Teg-3' . SUBSTIT T HE (lie) Preparation of an I-Q τ -I-Maleimide using Sulfo-SMCC: 5'-Teg-I-0 T -I-3'-Maleimide An aqueous solution containing 20 nmoles of 5'-Teg- I-Q-*-I-3'-NH 2 (prepared as in Example 2a) was diluted into phosphate buffed saline. Sulfo-SMCC (100 nmoles) in PBS was added and the resulting mixture was allowed to stand for 30 min. with occasional mixing at room temperature. The reaction mixture was diluted with phosphate buffed saline , added to a prewashed PD-10 column, and eluted with PBS to afford δ'-Teg-I-Q j -I-S'- Maleimide. This material was stored on ice until use. (lid) Preparation of an I-O^I-Maleimide. 5'- Maleimide-I-O j -I-3'-Teg is reacted with sulfo-SMCC in the same way as in lie to afford δ'-Maleimide-I-Q j -l- 3'Teg. (lie) Preparation of 3'-mercapto-I-O τ -I using SATA An aqueous solution containing 50 nmoles of 5'-Teg- I-Q t -I-3 ' -im 2 (prepared as in Example 2a) was diluted into PBS and 500 nmoles of SATA (in DMSO) was added. After mixing and standing at room temperature for 60 min, the reaction mixture was diluted with PBS, and eluted from a NAP-10 column with PBS to afford 5'-Teg-I- Q I -I-3'NH-CO-CH 2 -S-CO-CH 3 . The acetylthioacetylated oligonucleotide was deprotected by the addition of 30 μL of a pH 7.5 solution containing 100 mM sodium phosphate, 25 mM EDTA, 500 mM NH 2 OH. The reaction proceeded for two hourε at room temperature after which the material was again pasεed down a NAP-5 column by elution with PBS. SUBSTITUTE SHEET The final product (S'-Teg-I-Q j -I-S'NH-CO-CH 2 .SH) was used immediately. (llf) Preparation of 5'mercapto-I-O j -I using SATA H j N-δ ' -I-Q- t -I-S ' -Teg is reacted with SATA in the same way as in Example lie to afford 5'-HS-CH 2 -OC-N-I- Q- t -I-3'Teg. Example 12 (12a) Conjugation of 5'Teg-I-O j -I- 3 NHCfNH, * )CH H ? CH ? SH to Antibodv-Maleimide; ING-l-Maleimide-3'-S-I-O τ -I-5'-Teg A sample (108 nmoles) of 5'Teg-I-Q I -I-3'- NHC(NH 2 + ) CH 2 CH 2 CH 2 SH (prepared according to Example 11a) waε eluted off a NAP-25 column directly into a εolution of maleimide-derivatized ING-1 (18 nmoles) prepared according to Example 10a. After mixing, the reaction was allowed to proceed for 20 hours at 4°C. The reactants were then loaded into Centricon-100 ® concentration devices (Amicon) , and centrifuged at lOOOg for 25 minutes. The sample was resuspended in fresh PBS, and concentration by centrifugation was repeated a further 3 times until the ratio of optical densities at 260 nm and 280 nm was constant. The final product is ING-1- Maleimide-S-(CH 2 ) 3 -C(NH 2 ')NH-3'I-Q I -I-5'Teg. (12b) Conjugation of 5'-HSCHXH 2 (NH-. * . CNH-I-O τ -I-Teg- 3' to Antibodv-Maleimide, ING-l-Maleimide-5'- S-I-O τ -I-Teg-3' A sample (108 nmoles) of 5'-HSCH 2 CH 2 CH 2 (NH, * )CHN-I- Q j -I-Teg-3' (prepared according to Example lib) was SUBSTITUT HEE conjugated to 18 nmoles of. maleimide-derivatized ING-l aε in Example 12a to give ING-l-Maleimide-5'S-(CH 2 ) 3 - C(NH 2 + )NH-I-Q ϊ -I-3'Teg. (12c) Assays on the ING-1-Maleimide-S-I-O j -I con ugates The optical density of both ING-1-Maleimide-S- (CH 2 ) 3 .C(=NH)NH-I-Q I -I samples was examined in a spectrophotometer at 260 nm and 280 nm. The ratio of optical denεitieε at theεe two wavelengths was calculated and, by using known extinction coefficients for the antibody and for the oligonucleotide (approximate molecular weight 16500) at each of these wavelengths, the number of oligonucleotide molecules was estimated to be between 1 and 2 I-Q : -I per antibody. The concentrations of ING-l in the conjugate solutions were determined by the BioRad protein asεay uεing bovine immunoglobulin as the protein standard. These data agreed well with the antibody concentrations determined by examination of the optical density of the conjugate at 280 nm once it had been corrected for absorbance due to the conjugated I-Q j -I. Both these sets of data were further confirmed by subjecting the antibody-I-Q j -I conjugates to acid digestion and amino acid analysis. Antibody-I-Q x -I conjugates were examined for their ability to bind to antigens on the surface of a human tumor cell line to which the antibody had been raised. The immunoreactivity of the conjugates was compared by flow cytometry with a standard preparation of the antibody before being subjected to modification and conjugation to I-Q j -I. Target HT29 cells (a human adenocarcinoma cell line: ATTC) were grown to confluency in tisεue culture flaεkε using McCoy's media SUBSTITUTE SHEET supplemented with 10% fetal calf serum. The cells were harvested by scraping the flask walls with a cell εcraper. Cells from many separate flaskε were pooled, centrifuged to a pellet, resuspended at 5xl0 5 /m__. in a solution of ice-cold 50mM sodium phosphate with 150 mM sodium chloride buffer pH 7.4 (PBS) supplemented with 0.1% bovine serum albumin (Sigma) and 0.02% sodium azide (Flow buffer) . The cells were washed in this εa e buffer and then counted. An antibody εtandard curve waε conεtructed by diluting a εtock solution of ING-l with an irrelevant (non binding) , isotype-matched control antibody (human IgG to give a number of samples ranging in ING-l content from 10% to 100%. The standard curve was made in flow buffer so that each sample contained 1.0 μg antibody protein per mL. Samples from the εtandard curve and unknownε were then incubated with 5xl0 5 HT29 cellε at 4°C for 1 hour. After extenεive waεhing to remove unbound antibody, the cellε were reεuεpended in 100 μL flow buffer and incubated at 4°C for 1 hour with goat-anti-human antibody labeled with fluoreεcene iεothiocyanate (FITC) . After further waεhing in flow buffer the samples were analyzed by flow cytometry on a Coulter EPICS 753 flow cytometer. Fluorescene from FITC and propidium iodide (PI) was excited using the 488 nm emisεion line of an argon laεer. The output was set at 500 mw in light regulation mode. Single cells were identified by 90 degree and forward angle light scatter. Analysis windows were applied to these parameters to separate single cells from aggregates and cell debris. Fluorescence from FITC and propidium were separated with a 550nm long pass dichroic filter and collected through a 530 nm band pass filter (for FITC) , and a 635 nm band pass filter (for PI) . Light scatter parameterε were collected aε integrated pulses and fluorescence was collected as log SUBST T integrated pulεeε. Dead cellε were excluded from the aεεay by placing an analyεis window on cells negative for PI uptake. The mean fluorescence per sample (weighted average from 2500 cells) was calculated for each histogram. FITC calibration beads were analysed in each experiment to establish a fluorescence standard curve. The average fluorescence intenεity for each εample waε then expressed as the average FITC equivalents per cell. Immunoreactivity was calculated by comparing the average fluorescence intensity of the unknown sample with values from the standard curve. From the immunoreactivity assay, ING-1-Maleimide-S- (CH 2 ) 3 -C(NH 2 'f )NH-3'I-Q I -I 5'Teg was 67.8% as immunoreactive as the ING-l standard and ING-1- Maleimide-S-(CH 2 ) 3 -C(NH 2 * )NH-5'I-Q I -I 3'Teg was 81.5% aε immunoreactive. In a εeparate εet of experimentε, immunoreactivity of ING-1-I-Q j -I (3' conjugate) and ING- 1-I-Q j -I (5' conjugate) were determined to be 68% and 59% reεpectively. SUBSTITUTE SHEET Table 1 Preparation ING-1-Maleimide-S- (CH 2 ) 3 -C (NH 2 + ) NH-3 ' I-Q j -I 5 ' Teg ING-1-Maleimide-S- ( CH 2 ) 3 -C (NH 2 + ) NH-5 ' I-Q j -I 3 ' Teg ING-l antibody pre-conjugate Samples of these conjugates were also subjected to electrophoresiε on Novex 6% polyacrylamide gelε using SDS buffers in order to estimate their apparent molecular weight and the degree of heterogeneity of the preparation. Uεing εtandards of known molecular weight run on the same gel, a standard curve was constructed of the distance travelled (Rf) versus the log of the molecular weight. From this standard curve the relative molecular weights of the bandε aεεociated with each conjugate preparation were determined (see Table 1) . The SDS PAGE gels of INGl-I-Q j -I and ING-l antibody demonstrated that the molecular weight of the ING-1-I- Q j -I conjugates were higher than that of the antibody alone. SUBS (1 d) Conjugation of Teg-5'-I-0 I -I-3 '-Maleimide to ercaptoalkyl-Antibody 20 n oleε of Teg-5'-I—Q x -I-3 ' Maleimide (from Example lie) were reacted with Snmoleε of ercaptoalkyl- ING-l (from Example 10b) in PBS pH 7. After mixing the reaction waε allowed to continue at 4°C for 16 hourε to afford ING-l-NH-CO-CHj-S-Maleimide-S'-I-Q ϊ -I-S'Teg. The reaction mixture was loaded into a Centricon-100 ® concentration device and centrifuged at lOOOg for 25 minutes. The sample was resuspended in fresh PBS and concentration by centrifugation was repeated a further 3 times until the ratio of optical densities at 260 nm and 280 nm was constant. Maleimide-S'-I-Q j -I-S'Teg is reacted with mercaptoalkyl-ING-1 in the same way to afford ING-l-NH- CO-CH ^ -S-Maleimide-δ'-I-Q^I-S'Teg. (12e) Conjugation of Teg-δ'-I-O^I-S 'N-CO-CH ; _SH to Antibodv-Maleimide A 6 nmole sample of ING-1-Maleimide (from Example 10a) in PBS was reacted with 40 nmoles of Teg-δ'-I-Q j -I- 3'NH-CO-CH 2 _.SH (from Example lie) at 4°C for 16 hours. The reactantε were diluted with PBS and eluted in PBS from a PD-10 column to afford ING-l-Maleimide-S-CH 2 -OC- NH-S'-I-Qj-I-S'Teg. The product waε concentrated in a Centricon-300' device by centrifugation at lOOOg for 25 minutes. The sample was resuspended in fresh PBS and concentration by centrifugation was repeated a further 3 times until the ratio of optical densities at 260 n and 280 nm was constant. HS-CHj-OC-NH-δ'-I-Q ϊ -I-S'Teg is reacted with ING-1- Maleimide in the same way to afford ING-1-Maleimide-S- CH,-OC-NH-5'-I-Q I -I-3'Teg. SUBSTITUTE SHEET The optical denεity of theεe samples is examined in a spectrophotometer at 260 nm and 280 nm. The ratio of optical densitieε at theεe two wavelengths is calculated and, by using known extinction coefficientε for the antibody and for the oligonucleotide (approx molecular weight 16500) at each of these wavelengths, the number of oligonucleotide molecules per antibody is estimated. (12f) Conjugation of 5'-Trityl(S)-I-O τ -I-3'-NH-, to Antibodv-Maleimide A sample (10 nmoles) of 5'-Trityl(S) -I-Q τ -I-3'-NH 2 synthesized according to Example 2c, was diluted into PBS and a solution of silver nitrate in water was added to a final concentration of 85 mM. After vortexing the reaction was allowed to proceed at room temperature for 30 minutes. A precipitate formed which was centifuged to the bottom of the tube. The clear supernatant, which contains the oligonucleotide with a 5' terminal thiol group and without the 5' trityl group (HS-δ'-I-Q j -l-S'- NH 2 ) was kept at 4°C until use. A 6 nmole sample of ING-1-Maleimide (prepared essentially according to Example 10a) in PBS was reacted with 40 nmoleε of HS-δ'-I-Qj-I-S'-NH 2 at 4"C for 16 hourε. The reactants were diluted with PBS and eluted in 4 L from a pre-washed Econopac 106-DG column (BioRad) to afford ING-l-Maleimide-S-S'-I-Q- j -I-S'Teg. The product was concentrated in a Centricon-300" concentration device by centrifugation at lOOOg for 25 minutes. The sample waε resuspended in fresh PBS and concentration by centrifugation was repeated a further 3 times until the ratio of optical densities at 260 nm and 280 nm was constant. The optical density of these samples were examined in a spectrophotometer at 260 nm and 280 nm. The ratio of optical densities at these two wavelengths waε calculated and, by using known extinction coefficients for the antibody and for the oligonucleotide (approximate molecular weight=16500) at each of these wavelengths, the number of oligonucleotide molecules per antibody waε estimated. Example 13 (13a) Annealing of cl to ING-1-I-0 T -I A sample of ING-1-Maleimide-S-(CH 2 ) 3 -C(NH, * )NH-3 'I- Q j -I 5'Teg (24 pmoles), prepared according to Example 12a, and a sample of ING-1-Maleimide-S-(CH 2 ) 3 -C(NH 2 ÷ )NH- 5'1-Q j -I 3'Teg (25 pmoles) prepared according to Example 12d, were mixed in separate cuvettes with 16-fold excess of cl prepared according to Example 3c, in 50 mM PBS containing 1.0 mM EDTA and 100 mM NaCL, pH 7.2 at room temperature. The cuvettes were cooled to 20°C, loaded into a Cary 13 instrument , and analysed by UV light (260 nm) while the cuvette temperature was ramped up from 20°C to 80°C and then back down to 20°C at a rate of 0.5°C/min. Analysis of the data revealed that cl was able to hydridize to both the 3' and the 5' conjugates with ternary complex T m of 61.5°C and 60.4°C respectively. Similar experiments using TMT-cI-TMT (Example 8) demonstrated the annealing of TMT-cI-TMT to ING-1-Maleimide-S-(CH 2 ) 3 -C(NH 2 * )NH-3'I-Q : -I 5'Teg. (13b) Hybridization of ING-l-I-0-,-1 to 32 P labeled cl or 32 P labeled I in phosphate buffer and human serum An ING-l-Maleimide-S-(CH 2 ) 3 -C(NH 2 + )NH-3'I-Q I -I 5'Teg (4ul; lmg antibody/ml) solution (from Example 12a) and SUBSTITUTE SHEET 50 μL 32 P-labeled cl εolution (Example 4) were mixed with freεhly prepared human serum (200 μL) or PBS (200 μL; pH7.2) and incubated at 37°C for 2 hours. Aliquots were then subjected to SDS PAGE electrophoresis on an 8- 16% gel. The gels were examined by both autoradiography and on a phosphoimager system to show that 3 P-labeled cl waε able to hybridize with ING-1-I-Q- j -I in human serum as well as phosphate buffer solution (pH 7.2) . Samples were left at room temperature for 14 days in PBS and in serum and run on 8-16% SDS-PAGE gave similar patterns to each other and to gels incubated at 37°C for 2 hours suggesting that the conjugates were stable in serum for up to two weeks. 32 P-labeled I (Example 4) failed to εhow hybridization to ING-l-I-Q- t -I at any time as expected. (13c) Hybridization of 90 Y-TMT-cI-TMT- 90 Y to ING-l-I- An ING-l-Maleimide-S-(CH 2 ) 3 -C(NH 2 + )NH-3'I-Q I -I-5'Teg (4ul; lmg/ml) solution (from Example 12a) and 50 μL 90 Y- TMT-cI-TMT- 90 Y (Example 9) were mixed in PBS (200 ul; pH7.2) and incubated at 37°C for 60 minutes. Aliquots of these mixtures were then subjected to SDS PAGE electrophoresiε on an 8-16% gel. The gels were autoradiographed on a phosphoimager syεtem to εhow that 90 Y-TMT-cI-TMT- 90 Y waε able to hybridize with ING-I-I-Q* ! - I. (13d) Weεtern blot to detect TMT on TMT-cI-TMT hybridized with INGl-I-O- t -I. Sampleε of I-Q---I (Example 2a) or ING-1-Maleimide- 3'S-(CH 2 ) 3 .C(=NH 2 + )NH-I-Q I -I-5'Teg (Example 12a) were mixed with the oligonucleotide TMT conjugate (TMT-cI- SUBSTITUT HE TMT; Example 8) on ice for 10 min. Aliquots of these reaction mixtures were mixed with SDS buffer and loaded onto two duplicate 8-16% polyacrylamide gels. The gels were subjected to electrophoresis at a constant voltage for 2 hours. One gel was electroblotted onto nitrocellulose paper using CAPS buffer for 20 minutes according to the manufacturer's protocol (Hoefer semi- dry transfer method) . After washing thrice with a solution of 0.05% Tween 20 in PBS, the gel waε blocked with a εolution of 3% BSA in PBS at room temperature for 1 hour. Following further waεhing with Tween/PBS the gel waε overlaid with a εolution of a murine anti-TMT antibody (lOμg/ml in PBS/Tween) and left overnight at room temperature. The weεtern blot was developed using a goat anti-mouse IgG antibody conjugated to horseradiεh peroxidaεe (BioRad Weεtern blot kit) and peroxidaεe εubεtrate. The blot demonstrated that the TMTs of TMT- cI-TMT could be detected hybridized to bandε that contained either I-Q : -I or ING-1-I-Q j -I. The εecond gel waε εtained with ethidium bromide (5mg/ml in diεtilled water) . Examination of the εtained gel under UV light again demonstrated hybrization. Example 14 Preparation of Ab-TMT by direct conjugation (ING-1/TMT) TMT-NCS or a suitable derivative thereof can be conjugated to an antibody molecule to yield an antibody- TMT conjugate molecule that displays the ability to bind to a target antigen recognized by the antibody variable region. Such a conjugate molecule can be used to deliver metal ions that are chelated by the TMT moiety in order to localize and/or treat the lesion that is targeted by such an immunoconjugate. In one preferred SUBSTITUTE SHEET embodiment, the antibody is selected such that it has a broad reactivity with an antigen molecule expresεed on tumor cellε, thereby providing an antibody-TMT conjugate that can deliver metalε to the tumors for therapeutic or diagnostic purposeε. The chimeric antibody, ING-l, (International patent publication WO 90/02569) conεiεts of a murine variable region and a human immunoglobulin constant region. The antibody is produced by culturing a mouse myeloma cell line expressing the chimeric antibody essentially as described in the above- referenced publication. After purification, ING-l iε uεed at a concentration of 5.0 mg/mL in 50 mM sodium acetate and 150 mM εodium chloride buffer pH 5.6. The conjugation of ING-l to TMT-NCS iε achieved by firεt adding 1.0M carbonate, 150 mM εodium chloride buffer, pH 9.3 to ING-l until the antibody solution reaches a pH of 9.0. A sample of that ING-l solution, containing 5 mg of protein, is then pipetted into an acid washed, conical, glass reaction vial. A solution of TMT-NCS is prepared by dissolving 100 mg in 10 mL of 1.0 M carbonate, 150 mM sodium chloride buffer, pH9.0. The conjugation reaction is started by the addition of 96.5 μL of the TMT-NCS solution to the antibody to give a 4-fold molar excess of TMT-NCS over ING-l. The solution is stirred briefly to mix the reactants and then left in the dark at room temperature. After 16 hourε, the ING-1/TMT conjugate iε separated from unconjugated TMT by applying the reaction mixture to a PD10 chromatography column which has been pre-washed and equilibriated with 50 mM sodium acetate in 150 mM sodium chloride buffer, pH 5.6. The pure conjugate is eluted off the column with 2.5 mL of that same buffer. Example 15 Analytical tests on the ING-1/TMT conjugates. (15a) Analvsiε of Chelator to Antibody Ratio The protein concentrationε of ING-l in the conjugate εolutions were determined by the BioRad protein asεay using bovine immunoglobulin as the protein standard. In order to calculate the number of functional TMT molecules per antibody, ING-1/TMT is reacted with a solution of Europium chloride until saturation of the metal-binding capacity of the TMT occurs. A 0.375 mg aliquot of the ING-1/TMT in 2.5 ml in 0.05 M Tris HC1 buffer pH 7.5 is pipetted into a 5 ml quartz cuvette. A 20 μM Europium chloride (Europium chloride hexahydrate: Aldrich) solution in 0.05 M Tris HC1 buffer pH 7.5 is prepared. An aliquot (50 μL) of this Europium chloride solution iε added to the cuvette containing ING-1/TMT and the reεulting εolution slowly stirred on a magnetic stirrer at room temperature for 10 min uεing a εmall magnetic stir bar placed in the cuvette. The fluorescence of the metal-ING-1/TMT complex is determined in a Perkin Elmer LS 50 spectrofluorometer using an excitation wavelength of 340 nm (10 nm slit width) . The fluoreεcent emission iε monitored at 618 nm uεing a 10 nm εlit width and a 430 nm cutoff filter. The above procedure iε repeated and fluorescent readings are made after each addition. Aliquots of Europium chloride are added until the increase in fluorescence intensity is lesε than 5% of the preceding reading. A dilution correction iε applied to the fluoreεcence intenεity meaεured at each mole ratio, to compenεate for the change in volume of the teεt εolution. Since each SUBSTITUTE SHEET chelating site on the ING-1/TMT conjugate binds one Europium ion, and since Europium ion has to be in a chelate site for fluoreεcence to occur, thiε method allows the number of functional chelation siteε to be quantitated. Uεing thiε method, the ratio of TMT moleculeε per molecule of antibody is in the range 1: 1 to 2:1. (15b) ING-1/TMT Immunoreactivity assay by ELISA The antigen to which the antibody, ING-l, binds is prepared from LS174T or HT 29 cells (available from American Type Tissue Collection, ATTC) by scraping confluent monolayers of cells from the walls of culture flasks with a cell scraper. The cells from many flaskε are combined and a εample iε taken and counted to eεtimate the total number of cellε harveεted. At all timeε the cellε are kept on ice. Following centrifugation of the cellε at 1500 rpm for 10 minuteε at 4°C , the cellε are washed once in 25 mL ice-cold 50mM sodium phosphate buffer, pH 7.4 supplemented with 150 mM sodium chloride (PBS) , pelleted under the same conditions and transfered in 10 mL PBS to an ice-cold glass mortar. The cells are homogenized at 4°C using a motor-driven pestle and then centrifuged at 3000 x g for 5 minutes. The antigen-rich supernatant is removed from the other cell debris and subjected to further centrifugation at 100,000 x g for one hour at 4°C. The pellet (antigen fraction) from this final εtep iε εuεpended in 100 μL of PBS for every million cellε harveεted. Following an eεtimate of the protein concentration (BioRad BCA protein aεεay using bovine immunoglobulin as the protein standard) the antigen is stored at -20°C until uεe. SUBSTIT T H Each well of a 96-well Costar microtiter plates is coated with antigen by adding 100 μL/well of cell lyεate (10 μg/ml) prepared aε above. The microtiter plates are allowed to dry overnight in a 37°C incubator. After waεhing the plate five timeε with 0.05% Tween-20 (Sigma) they are blotted dry. The wellε of each plate are blocked by adding 125 μL/well of a 1% BSA (bovine serum albumin, Sigma A-7906) solution in PBS and incubated for 1 hour at room temperature. The plates are washed five times with 0.05% Tween-20. Samples (50 μL/well in duplicate) of ING-1/TMT conjugates and standard ING-l antibody solutionε are prepared at a range of concentrationε in 1% BSA in PBS. Biotinylated ING-l (1.0 μg/mL in 0.1% BSA) is added to each well (50μL/well) and the plates are then incubated for 2 hours at room temperature. Following five washeε with 0.05% Tween-20, the plates are blotted dry and incubated at room temperature for one hour with dilute (1:2000 in 0.1% BSA) streptavidin-alkaline phoεphataεe (Tago; #6567) . After a further five washes, color is developed in each well upon the addition of 100 μL per well of phosphatase subεtrate reagent (Sigma) . After one hour at room temperature, the color iε read using a 405 nm filter in a Titertek Multiscan microplate reader. When teεted by thiε procedure, the immunoconjugates of ING-l with TMT are found to have immunoreactivity comparable to native ING-l. (15c) ING-1/TMT Immunoreactivity Assay by Flow Cvtometrv Target HT29 cells are grown to confluency in tissue culture flasks using McCoy's media supplemented with 10% fetal calf serum. The cellε are harveεted by scraping the flask walls with a cell scraper. Cells from SUBSTITUTE SHEET many separate flasks are pooled, centrifuged to a pellet, resuεpended at 5xl0 5 /mL in a εolution of ice- cold 50mM εodium phoεphate with 150 mM sodium chloride buffer pH 7.4 (PBS) supplemented with 0.1% bovine serum albumin (Sigma) and 0.02% sodium azide (Flow buffer). The cells are washed in this same buffer and then counted. An antibody standard curve is constructed by diluting ING-l with an irrelevant (non binding) , isotype-matched control antibody (human IgG to give a number of sampleε ranging in ING-l content from 10% to 100%. The εtandard curve iε made in flow buffer εo that each sample contains 1.0 μg protein per L. Samples from the standard curve and unknowns are then incubated with 5xl0 5 HT29 cells at 4°C for 1 hour. After extensive waεhing to remove unbound antibody, the cells are resuspended in 100 μL flow buffer and incubated at 4°C for 1 hour with goat-anti-human antibody labeled with fluorescene iεothiocyanate (FITC) . After further washing in flow buffer the sampleε are analyzed by flow cytometry on a Coulter EPICS 753 flow cytometer. Fluorescene FITC and propidium iodide (PI) are excited using the 488 nm emission line of an argon laser. The output is set at 500 mw in light regulation mode. Single cells are identified by 90 degree and forward angle light scatter. Analysiε windowε are applied to these parameters to separate single cells from aggregates and cell debriε. Fluoreεcence from FITC and propidium are εeparated with a 550nm long paεε dichroic filter and collected through a 530 nm band pass filter (for FITC) , and a 635 nm band pasε filter (for PI) . Light scatter parameters are collected as integrated pulses and fluorescence iε collected aε log integrated pulεeε. Dead cellε are excluded from the aεεay by placing an analysis window on cells negative for PI uptake. The mean fluorescence per sample (weighted average from 2500 SUBSTITUTE SHEE cells) is calculated for each histogram. FITC calibration beads are analysed in each experiment to establish a standard curve. The average fluorescence intensity for each sample is then expressed as the average FITC equivalentε per cell. Immunoreactivity iε calculated by comparing the average fluorescence intensity of the unknown sample with values from the standard curve. Samples of ING-1/TMT have immunoreactivity values comparable to the native ING-l antibody by this method. (15d) Determination of aggregate formation by size- exclusion HPLC. A 30 cm x 7.5 mm TSK-G3000SW size-excluεion HPLC column (Supelco) fitted with a guard column of the εame material iε equilibrated with 12 column volumes of 10 mM sodium phosphate buffer pH 6.0 supplemented with 150 mM sodium chloride using a Waters 600E HPLC syεtem with a flow rate of 1.0 mL per minute at 400-600 PSI. A εample (25μL) of BioRad gel filtration protein εtandards iε injected on to the column. The retention time of each εtandard iε monitored by a Waterε 490 UV detector set at 280 nm. Following the recovery of the final εtandard from the column, it iε waεhed with a further 10 volumes of 10 mM εodium phosphate buffer pH 6.0 supplemented with 150 mM sodium chloride. Sampleε (50μL) of either native ING-l antibody or ING-1/TMT at 200 μg/mL are injected on to the column and their retention times recorded. From the areas of the retained peaks and the retention time, the amount of aggregated material in the ING-1/TMT sample is calculated. By this method the native ING-l antibody has a retention time of 9.1 minuteε. ING-1/TMT haε a major peak alεo at 9.1 minuteε but a minor peak, attributable SUBSTITUTE SHEET to aggregates, can εometimeε be εeen at 7.3 minutes. By comparison of the peak areas, the aggregate peak is less than 5% of the total. (15e) Radiolabeling of ING-1/TMT with 90 Y. A volume of radioactive Yttrium chloride ( 90 Y in 0.04M hydrochloric acid at a specific activity of >500 Ci/mg: Amersham-Mediphysics) is neutralized using two volumes of 0.5 M εodium acetate pH 6.0. The neutralized 90 Y (1.0 mCi) iε added to 1.0 mL of ING-1/TMT (1 mg/mL) in 50 mM sodium acetate buffer containing 150 mM sodium chloride at pH 5.6. The labelling is allowed to proceed for one hour and then the reaction mixture is loaded on to a PD-10 chromatography column which has been pre¬ washed and equilibrated in a buffer containing 50mM sodium phosphate with 150 mM sodium chloride pH 7.4 (PBS). The sample is eluted from the column with 1.5 mL of PBS. Fractions of radiolabeled ING-1/TMT (0.5 mL) are collected, aεεayed for radioactivity, and pooled. The labeling efficiency iε determined by removing 1.0 μL of the sample and spotting it on to a Gelman ITLC-SG εtrip. The strip is developed in a glasε beaker containing 0.1 M sodium citrate, pH 6.0 for a few minuteε until the solvent front has reached three- quaterε of the way to the top of the paper. The εtrip is inεerted into a Syεtem 200 Imaging Scanner (Bioεcan) which haε been optimized for 90 Y and is controlled by a Compaq 386/20e computer. In this εyεtem free 90 Y migrateε at the εolvent front while the ING-1/TMT ( 90 Y ) remains at the origin. Using this syεtem more than 98% of the total 90 Y radioactivity iε alwayε found associated with ING-1/TMT at the origin. SUBSTIT (15f) Labeling ING-1/TMT with fluoreεcent metals. Binding of lanthanides εuch as europium (3 + ) to chelating agents that contain an aromatic moiety held close to the co-ordination sphere can lead to "senεitized" fluoreεcence wherein light is absorbed through the aromatic εyεtem and the energy is transfered to the metal. The metal then produces emissions characterized by a very large Stokes shift and fluoreεcence lifetimes of up to several seconds. A 0.5 mg aliquot of the ING-1/TMT in 2.5 mL in 0.05 M Tris HCl buffer pH 7.5 was pipetted into a 4 mL conical reaction vial containing a εmall εtirring bar. A 250 μM europium chloride (europium chloride hexahydrate: Aldrich) solution in 0.05 M Tris HCl buffer pH 7.5 was prepared. An aliquot (50 μL) of this europium chloride solution waε added to the reaction vial containing ING-1/TMT, and the reεulting εolution waε εtirred very slowly on a magnetic stirrer at room temperature. The labelling was allowed to proceed for one hour and then the reaction mixture was loaded on to a PD-10 chromatography column which had been pre-washed and equilibrated in a buffer containing 10 mM εodium phoεphate and 150 mM sodium chloride at pH 6.0 (PBS). The sample was eluted from the column with 3.5 L of PBS. The fluorescence of a 50 μL sample of the metal-ING-1/TMT complex was determined in a Perkin Elmer LS 50 spectrofluoro eter uεing an excitation wavelength of 340 nm (10 nm slit width) . The fluorescent emisεion waε recorded at 618 nm uεing a 10 nm slit width and a 430 nm cutoff filter. Each functional chelating site on the ING-1/TMT conjugate binds one europium ion. Using this method, between 1 and 3 fluoreεcent europium ionε were bound per molecule of antibody. SUBSTITUTE SHEET Example 16 (16a) Flow Cvtometry of binding of fluorescent cl to . cells treated with ING-1-I-O j -I. ING-1-Maleimide-S-(CH 2 ) 3 -C(NH 2 + )NH-3'I-Q^I 5'Teg was prepared as in Example 12a. ING-1-Maleimide-S- (CH 2 ) 3 -C(NH 2 + )NH-5'I-Q I -I 3'Teg was prepared as in Example 12b. Fluorescently labeled CY5.18-I-Q T -I waε prepared as in Example 2d. Fluorescently labeled CY5.18-CI and CY5.18-I were prepared as in Example 3e. Flow cytometry was carried out esεentially aε described in 12c except that the fluorescent dye CY5.18 was used in place of FITC. HT-29 cells (0.5xl0 6 ) were incubated on ice for 30 min. with 1 μg each of the ING-1-I-Q. j -I samples. The cells were washed x2 with flow buffer and pelleted at 1400 rpm x 5 minutes between washes. Next the cells in each sample were incubated with 5 μg CY5.18-cI or CY5.18-I for 3 hours on ice. Some cells were incubated with CY5.18-cI and CY5.18-I alone. After extensive washing with flow buffer, the cellε were subjected to analyεiε on a fluorescence activated cell sorter. CY5- 18 calibration beads were analysed to establish a standard curve of relative fluorescence intensity versus CY5-18 concentration. The mean fluorescence per sample (weighted average from 2500 cells) was calculated for each histogram. The average fluoreεcence intenεity for each εample waε then expressed as the average CY5-18 equivalentε per cell. Identical experimentε were carried out in which the medium used for incubation of the cells with the components was 100% fetal calf serum in place of flow buffer. Preliminary experiments showed that the time taken to maximize hybridization of CY5-18-cI to ING-1- Maleimide-S-(CH 2 ) 3 -C(NH 2 * )NH-3'I-Q I -I 5'Teg on the εurface SUBSTITUTE SHEET of HT-29 cells was about 3 hours. In Table 2 there waε no difference between flow buffer and 100% FCS in the degree of hybridization in each preparation εuggeεting that the end-capped oligonucleotide strands were stable to nuclease digestion. The data suggest that large amounts of CY5.18-cI can hybridize to both conjugates bound to the surface of cells. There was relatively little non-specifc binding to the cells either by the fluorescent oligonucleotides themselveε or by hybridization of CY5-18-I-Q J -I to the conjugates. (16b) Binding of 5 " ) Y-TMT-cI-TMT- 90 Y to HT-29 cellε treated with ING-1-I-O j -I. TMT-cI-TMT waε radiolabeled to a specific activity of 0.1 μCi/pmole as described " in Example 9. ING-1- TMT 90 Y was prepared as in Example 15e. ING-1-Maleimide- S-(CH 2 ) 3 -C(NH 2 + )NH-3'I-Q I -I 5'Teg was prepared as in Example 12a. Three tubes each containing 1x10 s HT-29 SUBSTITUTE SHEET cells in DMEM medium supplemented with 10% fetal calf serum were prepared and kept at 4°C throughout. Using matched protein concentrationε in all tubeε to enεure that the amount of added antibody waε equal, the tubes were treated as εhown in Table 3 below. Both sample 1 and sample 3 show higher radioactivity associated with the cell pellet than does sample 2 (directly labeled ING-1-TMT 90 Y) suggesting that a delivery system consisting of 2 separate components can deliver more radioactivity than a directly labeled antibody. SUBSTITUTE SHEE (16c) Binding of Europium labeled ING-1/TMT and TMT- cI-TMT conjugates to HT-29 cells and HT-29 cells treated with ING-1-I-0--I. respectively The TMT-cI-TMT conjugate was labeled with Europium ions as described in Example 8a and ING-1/TMT was labeled with Europium ions as described in Example 15f. Standard curves were created for both conjugateε in concentration rangeε of 100 picomoleε/100 μl to 6 femtomoleε/100 μl. HT-29 cellε were grown to confluence in McCoys media containing 10% FCS and 50 μgs/ml of gentamyacin. The cells were washed with phosphate buffered saline, and 5 ml of Trypsin Versene was added. The HT-29 cells were then incubated at 37 * C in 5% C0 2 for 15 minutes, complete media (5 ml) was added, and the cells were removed and washed in PBS. The HT-29 cells were then blocked with 10 μg of sheared salmon sperm DNA per 10e δ cells at 4°C for 30 minutes, washed in PBS and used in the hybridization assay as follows. 5 x 10 5 HT- 29 cells were added to a 100 μl working dilution of Ing- 1/TMT or Ing-1-I-Q- r -I and the mixture was incubated for 30 minutes on ice. The cellε were washed twice in 2 ml of a wash buffer (PBS + 0.1% BSA + 0.01% NaN 3 ) at 1400 RPM for 5 minutes. A working dilution of TMT-cI-TMT (lOOμl) was then added to the appropriate tubes and the reaction mixture was incubated on ice for 3 hours followed by washing twice in 2 ml of wash buffer (PBS + 0.1% BSA + 0.01% NaN 3 ) at 1400 RPM for 5 minutes. Each binding εtudy waε done in triplicate. The cell εuεpensions after binding and hybridization aε above were kept at 4°C in test tubes until use (10 minutes to one hour) . The Europium fluorescence was measured in a Delfia 1232 time-resolved fluorimeter by aliquoting four lOOμl portions into εeparate wellε in a Coεtar EIA/RIA 96-well plate from each tube after SUBSTITUTE SHEET vortexing. The resultε were proceεεed aε described in Example 15f. The counts per secondε (cps) of the Eu- TMTcI-TMT-Eu treated cells were considered as background and were subtracted from the cps data from the hybridization experiments (Eu-TMT-cI-TMT-Eu/Ingl-I-Q j - I) . This result and the cps data from the Ing-1/TMT-Eu binding experiment were translated as picomoles of bound TMT oleculeε per cell from the individual standard curves created independently. The results are described in Table 4 below. The results are described as fluorescence counts per second (cpε) or number (#) of picomoleε per 10 5 cellε. ING-1/TMT-Eu and ING-conjugates: 0.25 μg or 1.65 picomoles; Eu-TMT-cI-TMT-Eu: 100 ng or 15 picomoles per 5 X10 5 cells. Table 4 The invention has been described in detail with particular reference to certain preferred embodiments thereof, but it will be understood that variations and modifications can be effected within the spirit and scope of the invention. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: Snow, Robert A. Groves, Eric S. Shearman, Clyde W. Saha, Ashis K. Sen, Arup Black, Christopher D. V. (ii) TITLE OF INVENTION: SEQUENTIAL TARGETING OF TUMOR SITES WITH OLIGONUCLEOTIDE CONJUGATES OF ANTIBODY AND COMPLEMEMTARY OLIGONUCLEOTIDE CONJUGATES OF CHELATED RADIONUCLIDES (iii) NUMBER OF SEQUENCES: 36 (iv) CORRESPONDENCE ADDRESS: (A) ADDRESSEE: Dressier, Goldsmith, Shore, Sutker & Milnamo , Ltd. (B) STREET: 180 North Stetson, Suite 4700 (C) CITY: Chicago (D) STATE: Illinois (E) COUNTRY: USA (F) ZIP: 60601 (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 #1.25 (vi) CURRENT APPLICATION DATA: (A) APPLICATION NUMBER: (B) FILING DATE: (C) CLASSIFICATION: (viii) ATTORNEY/AGENT INFORMATION: (A) NAME: Katz, Martin L. (B) REGISTRATION NUMBER: 25,011 (ix) TELECOMMUNICATION INFORMATION: (A) TELEPHONE: (312) 616-5400 (B) TELEFAX: (312) 616-5460 (2) INFORMATION FOR SEQ ID NO:l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:l: TTATGGACGG AG 12 (2) INFORMATION FOR SEQ ID NO:2: (i) SEQUENCE CHARACTERISTICS: SUBSTITUTE SHEET (A) LENGTH: 13 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (geno_nic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: TTATGGACGG AGA 13 (2) INFORMATION FOR SEQ ID NO:3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: TTATGGACGG AGAA 14 (2) INFORMATION FOR SEQ ID NO: : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO: : TTATGGACGG AGAAG 15 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5: TTATGGACGG AGAAGC 16 (2) INFORMATION FOR SEQ ID NO:6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single SUBSTITUTE SHEET (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6: TTATGGACGG AGAAGCT 17 (2) INFORMATION FOR SEQ ID NO:7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7: TTATGGACGG AGAAGCTA 18 (2) INFORMATION FOR SEQ ID NO:8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8: TTATGGACGG AGAAGCTAA 19 (2) INFORMATION FOR SEQ ID NO: : (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9: CGGAGAAGCT AA 12 (2) INFORMATION FOR SEQ ID NO:10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 13 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) SUBSTITUTE SHEET (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10: ACGGAGAAGC TAA 13 (2) INFORMATION FOR SEQ ID NO:11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (Xi) SEQUENCE DESCRIPTION: SEQ ID NO:11: GACGGAGAAG CTAA 14 (2) INFORMATION FOR SEQ ID NO:12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12: GGACGGAGAA GCTAA 15 (2) INFORMATION FOR SEQ ID NO:13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13: TGGACGGAGA AGCTAA 16 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14: SUBSTITUT ATGGACGGAG AAGCTAA 17 (2) INFORMATION FOR SEQ ID NO:15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15: TATGGACGGA GAAGCTAA 18 (2) INFORMATION FOR SEQ ID NO:16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16: TTAGCTTCTC CG 12 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 13 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17: TTAGCTTCTC CGT 13 (2) INFORMATION FOR SEQ ID NO:18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: TTAGCTTCTC CGTC 14 (2) INFORMATION FOR SEQ ID NO:19: SUBSTITUTE SHEET (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 15 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: TTAGCTTCTC CGTCC 15 (2) INFORMATION FOR SEQ ID NO:20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20: TTAGCTTCTC CGTCCA 16 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE666XTYPBB.A (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21: TTAGCTTCTC CGTCCAT 17 (2) INFORMATION FOR SEQ ID NO:22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: TTAGCTTCTC CGTCCATA 18 (2) INFORMATION FOR SEQ ID NO:23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 base pairs (B) TYPE: nucleic acid SUBST T (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: TTAGCTTCTC CGTCCATAA 19 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 12 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24: CTCCGTCCAT AA 12 (2) INFORMATION FOR SEQ ID NO:25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 13 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25: TCTCCGTCCA TAA 13 (2) INFORMATION FOR SEQ ID NO:26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26: TTCTCCGTCC ATAA 14 (2) INFORMATION FOR SEQ ID NO:27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: IS base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear SUBSTITUTE SHEET (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27: CTTCTCCGTC CATAA 15 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28: GCTTCTCCGT CCATAA 16 (2) INFORMATION FOR SEQ ID NO:29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29: AGCTTCTCCG TCCATAA 17 (2) INFORMATION FOR SEQ ID NO:30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30: TAGCTTCTCC GTCCATAA 18 (2) INFORMATION FOR SEQ ID NO:31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 50 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) SUBSTIT T (ix) FEATURE: (A) NAME/KEY: miεc_feature (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "N equals Teg-T." (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 50 (D) OTHER INFORMATION: /note« "N equals NH2-T." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: NCTTATGGAC GGAGAAGCTA AACTCTCTTA TGGACGGAGA AGCTAATCTN 50 (2) INFORMATION FOR SEQ ID NO:32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 45 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: TCTTATGGAC GGATCCGCTA ATCTTATGGA CGGATCCGCT AATCT 45 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 52 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: miβc feature (B) LOCATION: 1 ~ (D) OTHER INFORMATION: /note= "N equals NH2 or Teg." (ix) FEATURE: (A) NAME/KEY: miβc_feature (B) LOCATION: 51 (D) OTHER INFORMATION: /note* "N equals NH2 or Teg." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: NTCTTATGGA CGGAGAAGCT AAACTCTCTT ATGGACGGAG AAGCTAATCT NT 52 (2) INFORMATION FOR SEQ ID NO:34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 50 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) SUBSTITUTE SHEET ( ix ) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "N equals NH2-T." (ix) FEATURE: (A) NAME/KEY: misc feature (B) LOCATION: 50 ~ (D) OTHER INFORMATION: /note* "N equals Teg-T." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34: NCTTATGGAC GGAGAAGCTA AACTCTCTTA TGGACGGAGA AGCTAATCTN 50 (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: misc feature (B) LOCATION: 1 ~ (D) OTHER INFORMATION: /note* "N equals NH2-T." (ix) FEATURE: (A) NAME/KEY: misc_feature (B) LOCATION: 20 (D) OTHER INFORMATION: /note* "N equals NH2-T." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35: NTAGCTTCTC CGTCCATAAN 20 (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (ix) FEATURE: (A) NAME/KEY: miβc_feature (B) LOCATION: 1 (D) OTHER INFORMATION: /note* "N equals NH2-T." (ix) FEATURE: (A) NAME/KEY: misc feature (B) LOCATION: 20 "" (D) OTHER INFORMATION: /note* "N equals NH2-T." (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: NTAT arcΛ ΛβΛJ-RCTAA 20 SUBSTITUTE SHEE

Claims: We cl a im : 1. A non-radioactive targeting immunoreagent that comprises a tumor antigen recognizing moiety, one or more oligonucleotides comprised of non self-associating oligonucleotide sequences, and one or more linking groups. 2. A radioactive targeting immunoreagent that comprises an oligonucleotide comprised of an oligonucleotide sequence that is complementary in sequence to and capable of hybridization with one or more fragments of a non-self-associating oligonucleotide sequence, one or more chelating agents, one or more linking groups and one or more radionuclides. 3. A targeting immunoreagent that comprises moieties repreεented by the structure 1: Structure 1 wherein: z compriseε the reεidue of an immunoreactive protein; L and Lς, are independently a chemical bond or a linking group; I comprises an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains one or more members of a family of homologouε contiguouε sequences, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units, provided that contiguous sequences of six or more nucleotide units of said oligonucleotide do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure 1; Q-[ is a spacing group; a iε 0 to about 6; Ii compriεeε an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units, a contiguous sequence therein comprising a portion of I; E is an end capping group; and p is an integer from 1 to about 10. 4. A radioactive targeting reagent comprised of moieties represented in the structure 2: Structure 2 wherein : cl compriεeε an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains therein one or more members of a family of homologous contiguous sequenceε, the individual homologε of said family being SUBSTITUTE SHE comprised of from 12 to about 30 nucleotide units; the nucleotide sequenceε of said members of said family of homologous contiguous sequences are complementary to the nucleotide sequences of members of the set of oligonucleotides in a coadministerable targeting immunoreagent; and contiguous sequences of six or more nucleotide units of said complementary oligonucleotide do not hybridize with any other contiguous sequences of six or more contiguous nucleotide units anywhere in structure 2; Q cl is a spacing group; L. , L 2 , and L 3 are independently a chemical bond or a lin ing group; W. , W 2 , and W 3 are residues of chelating groups; Mj, M 2 and M 3 are comprised of elements with oxidation states equal to or greater than +1, at least one of which is a radionuclide; x, y, and z are independently zero or one provided that at least one of x, y, or z is one; and w and b are independently zero or an integer from 1 to about 4. 5. The reagent of claim 3 wherein a is an integer from one to about 6. 6. The reagent of claim 3 wherein Z iε the residue of an antibody or antibody fragment. 7. The reagent of claim 6 wherein the antibody iε selected from ING-l; B72.3; 9.2.27; D612; UJ13A; NRLU- 10; 7E11C5; CC49; TNT; PR1A3 ; B174 ; B43; and anti-HLB antibodies. S . The reagent of claim 3 wherein L or L- is a residue of a heterobifunctional cross-linking reagent. SUBSTIT T HE 9. The reagent of claim 8 wherein the heterobifunctional crosε-linking reagent iε εelected from t e group consisting of sulfosuccinimidyl 4-(N- maleimidomethyl) cyclohexane-1-carboxylate, sulfoεuccinimidyl (4-iodoacetyl)aminobenzoate, sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate, 2- Iminothiolane, and N-succinimidyl S-acetylthioacetate. 10. The reagent of claim 3 wherein L or L g is a residue of a modified nucleotide moiety compriεing a reactive functional group. 11. The reagent of claim 10 where the reactive functional group iε selected from the group conεiεting of amine groups and sulfhydryl groups. 12. The reagent of claim 3 wherein I and Ii are DNA. 13. The reagent of claim 3 wherein I has the sequence shown in SEQ ID NO: 8. 14. The reagent of claim 3 wherein Q τ is the residue of an oligonucleotide. 15. The reagent of claim 14 wherein the oligonucleotide has the sequence ACTCTC. 16. The reagent of claim 3 wherein E iε the residue of a modified nucleotide group resistant to exonuclease activity. 17. The reagent of claim 16 wherein the modified nucleotide compriεeε a poly(alkylene glycol) phoεphate dieεter. SUBSTIT T 18. The reagent of claim 17 wherein the diester is tetra (ethylene glycol) phoεphate dieεter. 19. The reagent of claim 4 wherein cl haε the εequence εhow in SEQ ID NO: 23. 20. The reagent of claim 4 wherein Q τ iε the reεidue of an oligonucleotide. 21. The reagent of claim 20 wherein the oligonucleotide haε the εequence ACTCTC. 22. The reagent of claim 4 wherein L α , L, or L 3 is the reεidue of a heterobifunctional croεε-linking reagent. 23. The reagent of claim 22 wherein the heterobifunctional croεε-linking reagent iε εelected from the group conεiεting of εulfosuccinimidyl 4-(N- maleimidomethyl)cyclohexane-1-carboxylate, sulfosuccinimidyl (4-iodoacetyl)aminobenzoate, εulfosuccinimidyl 4-(p-maleimidophenyl)butyrate, 2- Iminothiolane, and N-succinimidyl S-acetylthioacetate. 24. The reagent of claim 4 wherein L j , L, or L 3 iε the reεidue of a modified nucleotide moiety containing a reactive functional group. 25. The reagent of claim 24 wherein the reactive functional group iε εelected from the group conεiεting of amine groupε and sulfhydryl groups. 26. The reagent of claim 4 wherein W- , W, or 3 independently contains a polycarboxylic acid group. SUBSTITUTE SHEET 27. The reagent of claim 4 wherein W lf W 2 or 3 is independently selected from the group consisting of B4A, P4A, TMT, DCDTPA, PheMT, macroPheMT, and macroTMT. 28. The reagent of claim 4 wherein the radionuclide is selected from 212 Pb, 212 Bi, 90 Y, 177 Lu, 1B6 Re, 188 Re, 6 Cu, 99m Tc, lια In, and 87 Y. 29. A method of making a compound of the structure: Structure 1 wherein: Z compriseε the reεidue of an immunoreactive protein; L and L Q are independently a chemical bond or a linking group; I compriεeε an oligonucleotide comprised of a contiguous sequence of from 12 to about 50 nucleotide units wherein said contiguous sequence contains one or more members of a family of homologous contiguous sequences, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units, provided that contiguous sequences of six or more nucleotide units of εaid oligonucleotide do not hybridize with any other contiguouε sequenceε of six or more contiguous nucleotide units anywhere in structure l; Q- is a spacing group; a is an integer from 1 to about 6; SUBSTITUTE SHEE Ii compriεeε an oligonucleotide compriεed of a contiguous sequence of from 12 to about 50 nucleotide units, a contiguous sequence therein comprising a portion of I; E is an end capping group; and p is an integer from 1 to about 10; compriεing: (i) derivatizing Z with a precursor to a residue of L under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of Z-[L] p ; (ii) derivatizing a precursor to a residue of E with a precursor to a residue of L g under conditions and for a time period εufficient to form a covalent complex which iε a precurεor to a residue of L g -E; (iii) derivatizing a precursor to a residue of L g -E with a precursor to a residue of I under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of Ii-L g -E; (iv) derivatizing a precursor to a residue of Ii-L g -E with a precursor to a residue of Q τ under conditions and for a time period sufficient to form a covalent complex which iε a a precurεor to a reεidue of Q I -I i -L c -E; (v) derivatizing a precursor to a residue of Q^Ii-L^-E with a precursor to a residue of l i and then with a precursor to a residue of Q : under conditions and for a time period sufficient to form a covalent complex which iε a precurεor of a reεidue of [Q τ -Ii] a -I^-E; (vi) derivatizing a precurεor to a residue of [Q- j -Ii] a - L p -E with a precursor to a residue of I under conditions and for a time period εufficient to form a covalent complex which iε a precursor to a residue of I-[Q ι -Ii] a - (vii) derivatizing a precursor to a residue of I-[Q : - I i ] a -L 5 -E with a precursor to a reεidue of L under SUBSTITUTE SHEET conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of L- (viii) derivatizing a precursor to a residue of L-I- [Q.-Ii] a -L ς -E with a precursor to a residue of Z-[L] p under conditions and for a time period sufficient to form a covalent complex Z-fL-I-fQ- f -I ] a -L g -E] p. 30. A method of making a compound of the structure: Structure 2 wherein: cl compriseε an oligonucleotide comprised of a contiguous εequence of from 12 to about 50 nucleotide unitε wherein said contiguous sequence contains therein one or more members of a family of homologous contiguous sequenceε, the individual homologs of said family being comprised of from 12 to about 30 nucleotide units; where the nucleotide sequenceε of said members of εaid family of homologouε contiguous sequences are complementary to the nucleotide sequenceε of memberε of the set of oligonucleotideε in a coadministerable targeting immunoreagent; and contiguous sequences of six or more nucleotide units of said complementary oligonucleotide do not. hybridize with any other contiguous sequences of SUBSTI six or more contiguous nucleotide unitε anywhere in structure 2; Q CI is a spacing group; L-, -L 2 , and L 3 are independently a chemical bond or a linking group; W-, W 2 , and 3 are residues of chelating groups; M lf M 2 , and M 3 are comprised of elements with oxidation states greater than +1, at least one of which is a radionuclide; x, y, and z are independently zero or one provided that at leaεt one of x, y, or z iε one; and w and b are zero or an integer from 1 to about 4 ; compriεing: (i) derivatizing a precurεor to a reεidue of , with a precurεor to a reεidue of cl under conditionε and for a time period εufficient to form a covalent complex which iε a precurεor to a residue of L α -cl; (ii) derivatizing a precursor to a residue of Q CI with a precursor to a residue of L 3 under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of Q CI -[L 3 ] W ; (iii) derivatizing a precursor to a residue ot I^-cl with a precursor to a residue of Q CI -[L 3 ] W under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of L 1 -cI-{Q CI -[L 3 ] w }; (iv) derivatizing a precursor to a residue of L j -cI-{Q CI - [ L 3 ] w } with a precursor to a residue of L 2 under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of L.-cI-{Q CI -[L 3 ] tt -L 2 }; (v) derivatizing a precursor to a residue of L j -cI-(Q CI - [L 3 ] w - L 2 ) with a precursor to a residue of Q cι -[L 3 ] W and then with a precursor to a residue of L 2 under conditions and for a time period sufficient to form a SUBSTITUTE SHEET covalent complex which iε a precurεor to a reεidue of (vi) derivatizing a precursor to a residue of L 2 -cI-[ Qcl - L 2 ] b I [L 3 ]„ with a precursor to a residue of W , W 2 , and W 3 under conditions and for a time period sufficient to form a covalent complex which is a precursor to a residue of Wj-Lj-ci-t Qd-L 2 ] b -w 2 ; [ 3 - 3 ] w (vii) derivatizing a precursor to a residue of with a precursor to a residue of [M-,],., [M 2 ] 2 , and [M 3 ] y under conditions and for a time period sufficient to form a complex 31. The method of claim 29 wherein Z is an antibody or antibody fragment. 32. The antibody of claim 31 wherein the antibody is εelected from ING-l; B72.3; 9.2.27; D612 ; UJ13A; SUBSTITUTE SHEET NRLU-10; 7E11C5; CC49; TNT; PR1A3 ; B174 ; B43; and and anti-HLB antibodies. 33. The method of claim 29 wherein L or - iε a residue of a heterobifunctional cross-linking reagent. 34. The method of claim 33 wherein the heterobifunctional cross-linking reagent is selected from the group consiεting of εulfoεuccinimidyl 4-(N- maleimidomethyl)cyclohexane-1-carboxylate, εulfoεuccinimidyl (4-iodoacetyl)aminobenzoate, εulfoεuccinimidyl 4-(p-maleimidophenyl)butyrate, 2- Iminothiolane, and N-succinimidyl S-acetylthioacetate. 35. The method of claim 29 wherein L or Lg is a reεidue of a modified nucleotide moiety containing a reactive functional group. 36. The method of claim 35 wherein reactive functional group is selected from the group conεiεting of amine groupε and εulfhydryl groupε. 37. The method of claim 29 wherein I and I are DNA. 38. The method of claim 29 wherein I haε the εequence εhown in SEQ ID NO: 8. 39. The method of claim 29 wherein Q τ iε a residue of an oligonucleotide. 40. The method of claim 39 wherein the oligonucleotide has the sequence ACTCTC. SUBSTITUTE SHEET 41. The method of claim 29 wherein E iε a residue of a modified nucleotide reεistant to exonuclease activity. 42. The method of claim 41 wherein the modified nucleotide comprises a poly(alkylene glycol) phoεphate dieεter. 43. The method of claim 42 wherein the phosphate diester is tetra(ethylene glycol) phosphate diester. 44. The method of claim 30 wherein cl haε the εequence εhow in SEQ ID NO: 23. 45. The method of claim 29 wherein Q, iε a reεidue of an oligonucleotide. 46. The method of claim 45 wherein the oligonucleotide haε the sequence ACTCTC. 47. The method of claim 30 wherein L*. , L 2 or L 3 is a residue of a heterobifunctional cross-linking reagent. 48. The method of claim 47 wherein the heterobifunctional crosε-linking reagent iε εelected from the group consisting of sulfosuccinimidyl 4-(N- maleimidomethyl)cyclohexane-l-carboxylate, sulfosuccinimidyl (4-iodoacetyl) aminobenzoate, sulfosuccinimidyl 4-(p-maleimidophenyl)butyrate, 2- Iminothiolane, and N-succinimidyl S-acetylthioacetate. 49. The method of claim 30 wherein L, , L, or L 3 iε a residue of a modified nucleotide moiety containing a reactive functional group. SUBSTITUTE SHEE 50. The method of claim 49 wherein the reactive functional group is selected from the group consisting of amine groups and sulfhydryl groups. 5 51. The method of claim 30 wherein W- , W, or 3 contains a polycarboxylic acid group. 52. The method of claim 30 wherein W 2 , 2 or W 3 iε selected from the group conεiεting of residues of B4A, 10 P4A, TMT, DCDTPA, PheMT, macroPheMT, and macroTMT. 53. The method of claim 30 wherein one of K, , M 2 and M 3 is a radioactive metal ion isotope. 15 54. The method of claim 53 wherein the radioactive metal ion isotope is 90 Y. 55. A pharmaceutical composition comprising a compound of claim 3 disεolved or dispersed in a 20 pharmaceutically acceptable carrier. 56. A pharmaceutical composition compriεing a compound of claim 4 diεεolved or diεpersed in a pharmaceutically acceptable medium. 2. z> 57. A method of treating a tumor in a mammal compriεing administering to said mammal an effective dose of a non-radioactive targeting immunoreagent of claim 3 in a pharmaceutically acceptable medium, waiting 30 for a time period sufficient for εaid non-radioactive targeting immunoreagent to accumulate at the tumor εite in said mammal, and subεequently, administering an effective doεe of a radioactive targeting reagent of claim 4 in a pharmaceutically acceptable medium to εaid 35 mammal, waiting for a time period sufficient for εaid SUBSTITUTE SHEET radioactive targeting reagent to accumulate at the target εite, εaid target εite being the εaid non- radioactive targeting immunoreagent accumulated at said tumor εite in εaid mammal. 58. A method of diagnoεtic imaging in a mammal comprising administering to said mammal an imaging effective dose of a non-radioactive targeting immunoreagent of claim 3 in a pharmaceutically acceptable medium, waiting for a time period sufficient for said non-radioactive targeting immunoreagent to accumulate at the imaging site in said mammal , and subsequently, administering an imaging effective dose of a radioactive targeting reagent of claim 4 in a pharmaceutically acceptable medium to said mammal, waiting for a time period sufficient for said radioactive targeting reagent to accumulate at the target site, said target site being the said non- radioactive targeting immunoreagent accumulated at said imaging εite in said mammal and generating an image of said mammal. 59. The reagent of claim 3 wherein I is a double helix. 60. The reagent of claim 4 wherein cl is a double helix. SUBSTITUTE SHEET

Inventors: BLACK CHRISTOPHER D V, GROVES ERIC S, SAHA ASKIS K, SEN ARUP, SHEARMAN CLYDE W, SNOW ROBERT A, BLACK, CHRISTOPHER, D., V., GROVES, ERIC, S., SAHA, ASKIS, K., SEN, ARUP, SHEARMAN, CLYDE W., SNOW, ROBERT, A.
Applicants: STERLING WINTHROP INC, STERLING WINTHROP, INC.
Application year: 1993
Main classification: A61K39
IPC codes: A61K39, A61K47, A61K51, C07H21, C12N15, C12P21, G01N33