MICROORGANISM IDENTIFICATION

Abstract: The invention relates to a method and apparatus for characterising microorganisms. The invention uses matrix assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF-MS) spectral data for a range of known microorganisms. This spectral data is included in a database for spectral data of each of known microorganisms. A sample of an unidentified microorganism is prepared and then subject to analysis by MALDI-TOF-MS techniques to produce the spectral data of the unknown organism. This data is then compared; using suitable comparison means with the spectral data in the database. Similarity indicates the genus while identity indicates the specific microorganism. The apparatus for this analysis has the advantage that once set up and calibrated, it is relatively simple to use.

Description: Micro-organism Identification The present invention concerns the identification of micro-organisms, and more particularly to the differentiation between strains of bacteria. The accurate identification of micro-organisms is time- consuming and complicated. As antibiotic-resistant strains of bacteria emerge there is a burgeoning need for fast and accurate identification of strains so that appropriate methods to combat them can be derived. Differentiation between strains of a bacterium is a very difficult process. Attempts have been made to distinguish between strains by the use of typing methodologies based upon phenotypic properties. The anti-biogram method attempts to distinguish differing strains by means of the variation in antibiotic resistance of each strain. Phage- typing distinguishes bacteria by infection with a virus. However, while such methods can group bacteria according to a range of phenotypic properties, they are not very effective at distinguishing between genera or strains of bacteria which have almost identical genetic constitution. Bio-typing has been used to try to provide the required differentiation. Bio-typing investigates the differences in, for example, toxin production and trytophan dependence between different strains. Even this method frequently fails to identify strains, even where the strains are derived from different patients and where a nosocomial incident (cross-infection) has occurred. Tracing the source(s) of nosocomial incidents is vital in places, such as hospitals where such incidents can have severe health implications. Correct identification of the strain of organism is vital to the prevention of further infection and to the treatment of those infected. This is particularly important with the recent emergence of Methycillin Resistant Staphylococcus Aureus (MRSA), many strains of which cannot be phage-typed. The analysis of mycobacteria present particular problems. The pathogenic forms, such as Mycobacterium tuberculosis, are typically slow to grow and accurate identification by traditional methods can take many weeks. However, early identification of tuberculosis is vital in the management of the infection in patients. Chromatographic analysis of derivatised mycolic acids and cell wall lipids is the current method for identification of mycobacteria. However such a method requires relatively large amounts of culture for the extraction and derivatisation steps . Identification by mass spectrometry combined with pyrolysis (Py-MS), gas chromatography (GC-MS) and fast atom bombardment (FAB-MS) has been applied to many areas of microbiology. However these methods require complex sample preparation and extraction procedures prior to analysis. In addition they require experienced specialist instrument operators which renders the microbiologist dependent upon external laboratories where such facilities can be maintained. The use of Matrix-Assisted Laser Desorption Ionisation Mass Spectrometry (MALDI-TOF-MS) for characterisation of biopolymers has been reported by Hillenkamp et al in 1991 Anal. Chem. 63, 1193A-1202A. Cotter has reported time of flight mass spectrometry for the structural analysis of biological molecules in 1992 Anal. Chem. 64, 1027A-1039A. There is a need for a rapid, accurate method of differentiating between organisms, which method is capable of differentiating between different strains where the differences are normally difficult to characterise . The present applicants have found that MALDI-TOF-MS spectral data for micro-organisms is spectral data which is dependent on the molecular structure of the microorganisms concerned. In effect, therefore, such data is a 'finger print' for the organism concerned. As a microorganism transmogrifies, its MALDI-TOF-MS spectral data will change to reflect the change in structure/composition of the organism. According to one aspect of the present invention, therefore, there is provided a method of characterising micro-organisms characterised by:- providing a database of matrix assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF-MS) spectral data for a range of known micro-organisms. preparing a sample of unidentified micro-organisms and obtaining spectral data thereof by MALDI-TOF-MS and comparing using suitable comparison means the spectral data so obtained with database to identify a known micro-organism having the same or similar spectral data. In another aspect of the present invention there is provided apparatus for the screening of micro-organisms characterised in that the apparatus comprises :- spectrσ etric means for producing MALDI-TOF-MS spectral data of a sample organism database means containing MALDI-TOF-MS spectral data for a range of micro-organisms. The invention enables the identification of microorganisms quickly and without the labour intensive procedures hitherto employed. MALDI-TOF-MS is a high performance mass spectrometry tool, which may be operated by the non-specialist. Furthermore, the equipment required to perform the analysis is relatively inexpensive. The method and/or apparatus of the invention uses dried, solid samples which are easy to handle and store, and which are capable of analysis in accordance with the invention over a wide mass range. Prior to analysis, the micro-organism may be cultured by conventional means, for example, on a nutrient agar plate. The spectrometer samples may be prepared by taking microorganism cells e.g. from a culture medium and applying them directly • to a sample plate and a matrix added. Alternatively the cells may be admixed with the matrix prior to application so that cellular integrity may be retained . The dried matrix-sample mixture is bombarded with laser energy to create gas phase ionic species which are then pulsed into a flight tube. Both positive and negative ions may be generated and identified, The species may be identified by their mass to charge ratio (m/z), which can be identified with an accuracy of better than 0.1%. The m/z value of each spectral peak may be identified from the centrσid of the peak corresponding to the to the average molecular mass of the particular M+H ion (in the positive ion mode). In another aspect of the invention, the spectral data of the unidentified micro-organism is derived from a plurality of laser shots of the sample. Preferably the position of the laser energy impinging on the sample is varied, between shots, over the sample spot. In this way the effect of heterogeneities in the sample may be reduced by the summing of spectra from each shot and/or by the exclusion of data statistically deviant from reasonable values. This multiple shot technique may assist accurate, reproducible spectral data to be produced. Preferably spectra are taken from each and every tracking position over the sample area. A linear analyzer may be used to enhance the sensitivity of the data where limited amounts of sample are available . The inventors of the present application have discovered that MALDI-TOF-MS produces unique spectra for each microorganism allowing identification and differentiation between different genera of micro-organisms and even within different strains. The present invention is also useful for the analysis of mycobacteria, since the MALDI-TOF-MS method requires very small amounts of material, can be completed within minutes, and requires no derivatisation step. In another aspect of the present invention each spectra compiled in the database is recorded as a graphical representation of a plot of intensity against mass to charge ratio. The spectral data may be recorded in graphic, numerical or electronic format, in digital or analogue form. Preferably each spectra is recorded on the storage medium of a computer or computer network. The storage medium may be magnetic, such as floppy disk, tape or hard disk. Alternatively the storage medium may be optical, such as CD-ROM or laser-disc, in another embodiment the storage medium may be ROM microchips. Preferably each spectra is stored as a series of mass to charge ratios each corresponding to a centroid of the peaks of each spectrum. By storing the mass to charge ratios alone, the minimum data required to characterize and/or identify the micro-oranisms is stored. To aid the search and identification process, the data may be arranged in groups of data corresponding to the genus of each micro-organism, with sub-divisions corresponding to the strain of micro-organism. Where a spectrum analyzed is not capable of identification, means may be provided to add the spectrum to the database for future reference. Where the database is stored in the storage medium of a computer, software may be provided to carry out the screening of the unidentified spectral data. The software may carry out the steps of acquiring and storing the unidentified micro-organism's spectral data by manual input or, preferably directly from the spectrometer, comparing the unidentified spectral data with the pre-characterised spectral data in the database and indicating where a match is found, or otherwise. In one embodiment, the database may include spectral data grouped according to the genus of each micro-organism and the software may be carry out a search strategy which involves comparing spectral data first to identify the genus of the unidentified bacteria and then to identify the strain of bacteria. In this way the search process is optimized to improve efficiency where a large range of spectral data is stored in the database. The software may be capable of allowing for calibration errors in the unidentified spectral data, so that the trends (e.g. the difference in m/z value between adjacent peaks in the data) may be screened with the corresponding data in the database rather than the absolute values. The computer may be incorporated into the MALDI-TOF-MS Spectrometer so that data is transferred directly from the detector to the computer for analysis. Alternatively the computer may be spaced apart from the spectrometer and linked by remote means. The present invention also includes a database of microorganisms comprising spectral data of each micro-organism obtained by MALDI-TOF-MS analysis for use in the method, or in conjunction with the apparatus, of the invention. Following is a description by way of example only and with reference to the figures of the drawings of a method of carrying the present invention into effect. Figure 1 is a graphical representation of partial spectra obtained from samples including SA, CF and ECC2. Figure 2 is a graphical representation of a partial, positive ion, mass spectrum of Mycobacteriu smegmatis, in the m/z range = 500-1500 Da. Each of the listed peaks A-K represent α number of possible isobaric species of mycoserates . The relative peak heights are related to the number of isobaric species present at each molecular weight. The peak letter, molecular weight, and one of the possible isobaric structures are listed as follows: A 1 156 l ,.|l,.lll j lV-V l 4 B 1 170 l l 7 ll 17 lll,IV 2 V l4 C 1 184 l 1β ll 1β lll IV-V 14 D tt tH rβ ll 1β lll j lV.V 14 E 1212,I 17 I1 17 III 4 IV 2 V 14 F 1226,l lβ ll 1β lll 5 IV-V 1 G 1240 l 1 ll ιa lll 4 IV j V„ H 1254 l 17 ll 17 lll s IV-V 14 I 126B,I 17 II, 7 III 5 IV 2 V 15 J 1282 l 1β ll 1 B lll 5 IV 2 V 14 K 1296 l 1β ll ιe lll 5 IV 2 V 15 The molecular weights quoted above are for the mono- lsotropic, molecular ion (M). In the following example ten micro-organisms were analyzed to produce the MALDI-TOF-MS spectra. The organisms were:- The ten micro-organisms were each sub-cultured in duplicate onto separate nutrient agar plates. Each pair was incubated at 37 ~ C, for three and six days respectively . A single, visible colony of cells was regarded as sufficient material for analysis and was removed from the agar plate with a sterile loop. All samples were analyzed in a matrix of α-cyano-4-hydroxycinnamic acid (α-CHCA). The organisms were emulsified in 50-100 ul of the matrix, dissolved in a mixture of water, ethanol and acetonitrile (in the ratio 1:1:1). A 1 ul aliquot was then applied to the sample target for the laser and allowed to dry before analysis. The samples were then analyzed by MALDI on a KRATOS" Kompact MALDI III time of flight mass spectrometer. A nitrogen laser giving a 337 nm output of 3 ns pulse width was use to ionize the species. The laser fluence was set just above the threshold for ion production. The mass spectrometer was used in the positive ion detection mode using an acceleration voltage of +20 KV. Samples were analyzed by using both the linear and reflectron analyzers. The detector used consisted of a discrete dynode combine with an electron multiplier. A spectrum was produced for each pair of each micro- organism. Figure 1 shows the spectra of SA, CF and ECC2. There is a clear difference in the distribution of the peaks corresponding to ionic species from each microorganism. The spectra produced constitute a unique " ingerprint" which allows identification of icro- organisms ihter-species and inter-strain. The similarities in the features of each spectra allow the spectra to be grouped according to genus of micro- organis , Figure 2 shows the positive ion spectrum of Mycobacterium smegmatis which shows an interesting pattern of peaks between 1150-1300 Da. Previous analyses of cell wall extracts from this organism suggest that these peaks are either mycolic acids or related lipids within the cell wall. The data probably relates to dimycocerosates of the phthiocerol family. These are non-covalently bound and are probably extracted into the matrix during the admixture process. The proposed structure of the mycoserates are shown in the legend of figure 2. Table 1 lists subsets of m/z values for all the micro- organisms examined. The m/z ratios of the spectral peaks of the four cultures of E.COLI bacteria are shown in the first column of table 1 for the m/z range 1058-1852 Da. The figure in bold type are peaks present in all the tested strains of the genus E.COLI (1102, 1231, 1316, 1360, 1489 and 1618). Thus the genus may be identified by the presence of these peaks. The strain may be identified by the absence or presence of further peaks. For example ECN has a peak at 1852 which differentiates from ECC1 and ECC2. ECK also has a peak at 1852, but may be differentiated by the additional peaks at 1058 and 1187. Different species which are closely related to E.COLI are CF and KA. These have peaks 1102, 1231, 1360, 1489, 1618 in common with E.Coli, but may be differentiated therefrom by the absence of the peak at 1316 which is characteristic of E.Coli. CF and KA may be distinguished by the absence or presence of the peaks at 750, 751, 1314, 1445, 1480, 1575, 1582 and 1876. The spectra produced, expressed in terms of mass to charge ratio of peaks in the spectra, provide a "fingerprint" for each micro-organism. The fingerprint is sensitive to both inter-genus differences, and inter- strain differences. A database of these fingerprints provides a method of rapidly screening samples to identify the strain of micro-organism present. This has particular application in the field of medical microbiology, -but will find application in many other fields where the precise and rapid identification of micro-organisms is required. A database of MALDI-TOF-MS spectral data was prepared in respect of the following organisms :- E.coli 8110 E.coli 10214 K12 lambda K.aerogenes 8172 C.freundii 9750 S.aureus Bl S.aureus B2 S.aureus methicillin resistant (MRSA) 01/270030 S.aureus (MRSA) 02/270238 *C. difficile NCTC 11204 *C. difficile NCTC 11206 *C . perfringens *C.tetani M.malmoense 0036c M. avium-intracellulare-scrofulaceum 0352c M. avium-intracellulare 0589c M.xenopi 0545c Nesseria species M96255506 Nesseria species M96255510 Nesseria species M97251540 Nesseria species M97251545 Nesseria species M97251682 The spectral data obtained is as follows :- ■ Λl Ir.-.acz Cell MALDI KRATOS DB Set E.coli 8110 Data: stdOOOl.l 24 Jul 97 16:26 Cal : -july97v3 24 Jul 97 11:05 Kraπos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE €> 1000 (bin 56) y.ass %Area %Total Apex(mV) Flags -48 Intact Cell MALDI KRATOS DB Set E.coli 8110 Data: εtdCOOl.l 24 Jul 97 16:26 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) Intact Cell MALDI KKATOS DB Set E.coli 10214 K121am Data: std0003.3 24 Jul 97 16:28 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) z Intact Cell MALDI KRATOS DB Set E.coli 10214 K121am Data: std0003.3 24 Jul 97 16:28 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1 : « ■ Linear High Power: 60, tDE @ 1000 (bin 56) Z-i Intact Cell MALDI KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 60, tDE θ 1000 (bin 56) Zl KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: Duly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE θ 1000 (bin 56) Intact Cell MALDI KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE ® 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 2388.8 1.10 0.24 0.43 2402.2 0.67 0.15 0.43 I Intact Cell MALDI KRATOS DB Set C.freundii 9750 Data: std0015.15 24 Jul 97 16:42 Cal: :uly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (b n 56) εs Intact Cell MALDI KRATOS DB Set Cfreundii 9750 Data: std0015.15 24 Jul 97 16:42 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) Zl Intact Cell MALDI KRATOS DB Set C.freundii 9750 Data : std0015.15 24 Jul 97 16:42 Cal: ;ιuly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 2222. 2. 39 0.29 0.34 2240. 1.03 0.13 0.25 2256. 9 1.61 0.20 0.34 2318.7 4.11 0.50 0.37 2370.4 0.88 0.11 0. .25 2414.7 3.11 0. 38 0. .28 2478.2 1.34 0.16 0. .40 U 2492.3 3.51 0.43 0. ,61 TJ 2542.1 2.85 0.35 0.34 2596.1 2.24 0.27 0.31 2682.0 1.48 0.18 0.25 2708.0 1.87 0.23 0.28 2733.6 1.77 0.22 0.28 2782.7 1.00 0.12 0.21 2816.6 3.42 0.42 0.31 2874.5 1.64 0. 20 0.34 2899.9 2.48 0.30 0.28 - 2946.3 0.82 0.10 0.25 2991.1 1.52 0.19 0.25 3087.4 2, 11 0.26 0.28 3162. .9 5.73 0.70 0.89 3188..4 2 41 0 29 0.37 3257..0 1 81 0 22 0.34 3400..2 100 00 12 18 19.61 U 3423..9 22 22 2 71 3.52 U 3501..7 0.45 0.05 0.21 3538..7 2.18 0.26 0.25 3586..0 2.16 26 0.25 3627.4 1.16 14 0.28 3769.8 1.49 18 0.28 3824.5 0.63 08 0.25 3874.8 5.38 66 0.55 3974 1.63 0.20 0.21 4769 .3 1.88 0.23 0.43 5451.2 42.34 5.16 7.35 7739.5 3.92 0.48 0.61 Zr S. aureus isolates Nutrient Agar Bl Data: aOOOl.l 17 Jul 97 17:18 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) ags Z8 S. aureus isolates Nutrient Agar Bl Da t a : aOOOl.l 17 Jul 97 17:18 Cal: test 25 Ap r 97 li -^ Kr Ma.to»«s Kompa»c_t_.MA„_LDI 2: V v 5 o* 2 **•• 1 • + ri - u v . Mass. i -' ■• + Lin n .ear High Power: 70, tDE @ %Area %Total Apex(mV) Flags' 1000 (bin 56) 1388.1 0.91 0.19 0.46 1426.4 2.40 0.51 0.64 1458.0 3.16 0.68 0.61 1473.2 1.29 0.28 0.55 1518.6 3.02 0.64 0.43 1534.8 1.69 0.36 0.55 1554.9 1.75 0.37 0.52 Tj 1569.1 1.68 0.36 0.52 U 1827.3 1.01 0.22 0.55 1841.9 1.00 0.21 0.46 2000.7 2.13 0.45 0.43 2100.5 2.59 0.55 0.43 2387.0 1.59 0.34 0.46 2516.3 1.38 0.29 0.49 2814.5 3.14 0.67 0.46 S . aureus isolates Nutrient Agar B2 Data: a0003.3 17 Jul 97 17:22 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 70, tDE @ 1000 (bin 56) o : test 25 Apr 97 11:25 inear High Power: 70, tDE @ 1000 (bin 56) ags 3- S. aureus isolates Nutrient Agar B2 Data: a0003.3 17 Jul 97 17:22 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 VS .2.1 : * Linear High Power: 70, tDE @ 1000 (bin 56) tDE @ 1000 (Din 56) Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97.11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) ags Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) 35 Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power. 70, tDE @ 1000 ( D in 56) Intact Cell MALDI MRSA 02/270238 Data: a0002.3 3 Jul 97 17:53 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 70, tDE @ 1000 (bin 56) 3> Intact Cell MALDI c. diff NCTC 11204 Data: diff0032.12 18 Jun 97 15:11 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1: - Linear High Power: 90, tDE @ 1000 (bin 56) 3? Intact Cell MALDI c. diff NCTC 11206 Data: dιff0033.13 18 Jun 97 15:12 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1: - Linear High Power: 90, tDE @ 1000 (bin 56) 33 Intact Cell MALDI c. diff PERF Data: dιff0037.17 18 Jun 97 15:19 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1 : - Linear High Power: 95, tDE @ 1000 (bin 56) Intact Cell MALDI c . di f f TETANI Data: dιff0039.19 18 Jun 97 15:22 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1 : - Linear High Power: 95, tDE @ 1000 (bin 561 Ui Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 3819 (bin 109) Mass %Area %Total Apex( V) Flags 792.6 0.80 0.05 0.31 838.0 0.92 0.06 0.34 850.1 1.78 0.11 0.67 U 863.1 24.18 1.44 6.00 U 877.6 4.43 0.26 1.38 U 883.8 6.54 0.39 1.69 U 896.5 4.01 0.24 1.16 904.4 3.32 0.20 1.32 U 912.4 12.09 0.72 3.16 TJ 922.7 4.80 0.29 1.26 U 938.9 2.23 0.13 0.43 U 950.2 21.29 1.27 4.50 U 964.6 3.03 0.18 0.80 U 978.4 45.32 2.71 11.46 U 993.0 9.20 0.55 2.51 TJ 1006.4 100.00 5.97 24.97 U 1020.9 5.73 0.34 1.90 U 1034.8 36.44 2.18 11.46 U 1040.0 34.03 2.03 10.75 U 1051.0 8.36 0.50 1.84 U 1067.6 58.51 3.49 13.20 U 1080.0 8.21 0.49 1.93 U 1094.7 29.85 1.78 7.44 TJ 1108.5 3.80 0.53 2.18 TJ 1122.1 12.20 0.73 3.46 U 1136.4 13.15 0.79 2.45 U 1149.8 5.72 0.34 1.75 U 1166.6 10.02 0.60 2.11 TJ 1180.2 4.50 0.27 1.16 U 1195.2 7.04 0.42 1.16 U 1205.0 4.06 0.24 0.89 TJ 1218.5 6.06 0.36 1.59 U 1231.7 7.10 0.42 1.59 U 1245.7 8.62 0.51 2.27 U 1259.8 13.71 0.82 2.91 U 1277.3 12.19 0.73 2.82 TJ 1288.5 10.42 0.62 2.54 TJ 1304.1 34.21 2.04 5.94 TJ 1318.5 13.90 0.83 3.65 U 1332.6 13.05 0.78 2.91 TJ 1346.5 18.15 1.08 5.02 TJ 1361.1 15.28 0.91 3.80 TJ 1373.7 7.52 0.45 1.96 U 1388.4 10.30 0.62 2.21 U 1404.7 3.75 0.22 1.38 U 1416.8 6.92 0.41 1.84 TJ 1425.3 3.95 0.24 1.38 U 1432.5 4.51 0.27 1.10 U 1445.8 8.30 0.50 1.53 U 1458.0 4.88 0.29 1.16 TJ 1475.8 15.08 0.90 3.52 U 1493.6 16.57 0.99 1.87 U 1532.3 20.83 1.24 1.50 TJ 1555.7 5.54 0.33 1.38 U 1573.2 17.27 1.03 3.40 TJ 1591.3 14.10 0.84 3.74 U 1598.1 19.52 1.17 2.70 TJ m Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75. tDE @ 3819 (bin 109) Mass %Area %Total Apex(mV) Flags 1626.2 4.31 0.26 1.44 U 1646.5 16.75 1.00 2.54 U 1671.9 8.90 0.53 1.62 U 1681.6 20.03 1.20 3.49 U 1703.8 15.34 0.92 3.40 U 1715.9 1.37 0.08 0.74 U 1732.0 11.64 0.70 2.76 U 1741.1 12.82 0.77 3.03 U 1755.3 16.73 1.00 5.09 U 1760.9 13.23 0.79 3.71 U 1773.2 2.20 0.13 0.83 U 1787.1 5.27 0.32 1.26 U 1802.0 5.91 0.35 1.47 TJ 1817.7 98.66 5.89 18.72 U 1839.9 10.54 0.63 2.57 U 1866.8 7.46 0.45 1.19 U 1880.3 2.03 0.12 0.58 U 1889.7 10.39 0.62 2.70 U 1907.5 3.85 0.23 1.19 U 1917.0 7.30 0.44 1.72 U 1929.0 6.41 0.38 1.78 U 1935.2 3.23 0.19 1.10 U 1949.8 5.92 0.35 1.19 U 1963.2 5.03 0.30 1.26 U 1973.7 30.14 1.80 5.06 U 1988.9 23.53 1.41 5.33 U 2004.1 18.22 1.09 3.92 U 2018.9 18.35 1.10 3.68 U 2031.3 3.02 0.18 1.23 U 2044.5 6.76 0.40 1.78 TJ 2063.8 5.01 0.30 0.92 U 2080.1 7.17 0.43 1.65 U 2091.4 6.06 0.36 1.29 U 2104.8 2.85 0.17 1.16 U 2110.9 4.51 0.27 1.50 U 2120.9 7.32 0.44 1.87 U 2133.1 7.95 0.47 1.72 U 2153.7 7.88 0.47 2.18 U 2160.3 6.64 0.40 1.65 U 2171.3 3.65 0.22 1.23 U 2182.7 4.34 0.26 1.19 U 2201.3 11.27 0.67 2.24 U 2214.2 11.61 0.69 2.02 U 2234.8 11.92 0.71 2.94 U 2254.5 19.55 1.17 4.60 U 2271.6 15.04 0.90 3.46 U 2286.9 4.88 0.29 1.32 U 2305.0 26.39 1.58 6.65 U 2319.1 14.10 0.84 2.82 U 2336.5 4.34 0.26 1.35 U 2350.7 3.33 0.20 0.89 U 2369.1 10.27 0.61 1.50 U 2417.6 9.35 0.56 1.19 U 2433.0 14.51 0.87 3.00 U 2443.8 6.35 0.38 1.47 U 2461.1 22.88 1.37 4.53 U 2488.8 2.82 0.17 0.67 U U Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, tDE @ 3819 (bin 109) Vi Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16:27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56] Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16:27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) t Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16:27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) u tj iJ fc i i ιt> j i U (» ) o m w uι m M iθ ( Λ j ιo ιti W w o o c3 o o o o o o o o o o o o o o o o o o o o o μ-* ι-* to o o t-' θ σ o o o o o o o o o o o o M io w ι to u uι oi M i μ μ Λ t j ω w ι o μ M ^ m * iii ( B θ vi to M Ui fc w μ » w M μ o o μ ιπ u ιo t (_ ω ιo θ Φ μ øι ιo uι ιo tJ ι ι. aι () ι ^ uι ^ « o «l Ni uι θ Ln ιιi vi <k i- ω o i μ ι » ^ sj ιn ►" • l^ h* I-* μ* h-' M μ J O O I J O O O O θ μ J tO o ^ n ιl>. U J ι LO μ^ μ J O O O O O O O O O tD μ ) μ μ i to fc o ω ω w w m vo o ^ tD ifl uι μ μ Λ m oo J u a) sj ^ . vj w θ NJ ui (D ^ m ui fc ω u) J o t ui oi ui μ ^ i H t cti ti o io i H Ui totD to o ^ ui σi u μ ^ tn ixi tn H ui uKk A w βiσi μ Vito o u GGGGGGGGGGGGGGGGGGCGG G GGG GGGGGG GG GGGGG U9 Intact Cell MALDI Myco Cardiff 33/0589c Data: CAR0086.6 3 Jun 97 16: 37 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2 , 1: + Linear High Power: 75, tDE @ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 1667.3 5.64 0.44 2.70 1680.8 14.52 1.14 6.80 1696.0 15.33 1.20 6.28 1714.7 36.25 2.84 14.86 U 1729.2 98.68 7.72 34.93 U 1744.2 57.05 4.46 23.65 U 1757.3 61.17 4.78 26.87 U 1771.6 100.00 7.82 42.74 U 1786.3 80.92 6.33 34.87 TJ 1800 50.42 3.94 23.19 U 1814 36.86 2.88 16.64 TJ 1828 30.88 2.42 14.43 U 1842 10.60 0.83 5.27 1857.0 2.74 0.21 1.41 1871.7 2 22 0.17 1.26 1887.7 2 16 0.17 0.74 U 1904.6 3 73 0.29 1.16 U 1914 4 05 0.32 1.38 U 192 2.46 0.19 0.95 U 1941 5 97 47 1.32 U 1961 5 53 43 1.87 TJ 1975 6 85 54 2.18 TJ 1989.7 11 09 87 3.40 U 2001.5 6 43 50 2.27 U 2018.9 5 .65 44 2.11 U 2032. 6 .17 0.48 1.50 U 2062. 3 .43 0.27 0.98 U 2073. 3 .62 0.28 1.19 U 2088. 1 .63 0.13 0.89 U 2096. 1 .64 0.13 0.86 TJ 2108. 4 .11 0.32 1.32 U 2119.1 7 .04 0.55 2.73 U 2133.1 7 .93 0.62 2.48 U 2163.8 8.24 0. 64 1.75 U 2174.8 5.87 0.46 1.35 TJ 2194.7 77 0.06 0.58 U 2204.9 96 0.31 0.95 U 2223.6 81 0.22 0.83 TJ 2234.8 96 0.15 0.89 U 2245.5 07 0.32 .07 U 2259.0 26 0.25 .44 U 2269.8 26 0.10 .77 U 2285.1 20 0.25 .95 TJ 2291.4 24 0.10 80 U 2299.6 10 0.16 10 u 2306.9 32 0.18 38 U 2313.7 44 0.19 95 U 2328.3 2.38 0.19 0.89 U 2342.4 4.14 0, 32 1.19 U 2361.2 87 0.30 1.59 U 2371.8 83 0.77 2.88 U 2388.9 ,20 0 ,25 1.13 U 2399.5 .40 0.11 0.83 U 2419.0 .38 0.26 0.80 TJ 2433.9 .72 0.21 1.10 u 2461.6 .96 0.47 1.19 U 2479.4 .27 0.26 1.29 U V Intact Cell MALDI Myco Cardiff 33/0589c Data: CAR0086.6 3 Jun 97 16:37 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) Intact Cell MALDI Myco Cardiff 63/0545c Data: CAR0146.6 4 Jun 97 14:12 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) Intact Cell MALDI Myco Cardiff 63/0545c Data: CAR0146.6 4 Jun 97 14:12 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, tDE @ 1000 (bin 56) S Intact Cell MALDI Myco Cardiff 63/0545c Data: CAR0146.6 4 Jun 97 14:12 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, t DE Q 1000 [bin 56) Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25- Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 60, tDE @ 1000 (bin 56) St, Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power : 60, tDE @ 1000 ( bin 56 ) Mass %Area %Total Apex(mV) Flags 2504.4 0.52 0.07 0.12 2522.5 0.22 0.03 0.09 2533.4 0.46 0.06 0.12 2543.4 2.80 0.36 0.18 2552.5 0.77 0.10 0.12 2564.9 2.00 0.26 0.18 2575.0 0.95 0.12 0.12 2585.6 0.58 0.08 0.12 2593.3 1.14 0.15 0.12 2601.4 1.91 0.25 0.18 2627.1 0.89 0.11 0.12 2639.7 7.75 1.00 0.52 2652.8 0.31 0.04 0.09 2660.1 0.58 0.08 0.12 2678.7 2.43 0.31 0.18 2685.6 4.00 0.51 0.28 2696.4 0.77 0.10 0.12 2702.3 3.32 0.43 0.21 2720.0 0.68 0.09 0.12 2740.8 0.18 0.02 0.09 2746.7 0.71 0.09 0.12 2757.6 1.29 0.17 0.18 2771.5 2.52 0.32 0.21 2779.5 0.18 0.02 0.09 2784.5 0.31 0.04 0.09 2790.0 0.86 0.11 0.12 2794.5 1.14 0.15 0.12 2808.0 2.28 0.29 0.21 2829.1 0.83 0.11 0.15 2833.6 1.20 0.15 0.18 2848.7 3.63 0.47 0.28 2858.9 10.80 1.39 0.37 2900.5 5.42 0.70 0.25 2918.8 3.66 0.47 0.31 2928.6 1.14 0.15 0.15 2942.9 1.23 0.16 0.18 2951.1 1.17 0.15 0.15 2958.8 0.77 0.10 0.12 2962.9 0.31 0.04 0.09 2972.7 2.28 0.29 0.15 2984.6 1.88 0.24 0.25 2999.6 0.74 0.09 0.12 3005.8 0.31 0.04 0.09 3011.5 0.74 0.09 0.15 3019.3 1.45 0.19 0.12 3033.9 0.58 0.08 0.12 3037.5 0.98 0.13 0.15 3047.5 1.42 0.18 0.18 3072.6 2.74 0.35 0.18 3087.8 1.08 0.14 0.09 3096.2 0.83 0.11 0.12 3101.5 0.46 0.06 0.12 3109.9 0.31 0.04 0.09 3115.2 0.34 0.04 0.09 3120.5 1.11 0.14 0.18 3142.2 5.14 0.66 0.21 3146.9 0.22 0.03 0.09 Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 60, tDE 9 1000 (bin 56) Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) lags Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56 ) S8 Intact Cell MALDI Meningo 5/M96255510 Data: a0009.9 3 Jul 97 15:03 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 58, tDE @ 1000 (bin 56) 55 Intact Cell MALDI Meningo 5/M96255510 Data: a0009.9 3 Jul 97 15:03 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 58, tDE t? 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 985. 2.36 0.24 52 989. 1 0.46 0.05 31 995. 1 1.51 0.15 31 1003. 1 0.90 0.09 28 1018. 2 5.30 0.54 70 1031. 5 0.90 0.09 37 1043. 1 2.74 0.28 0.49 TJ 1047. 9 0.97 10 0.40 U 1056. 5 4.84 50 0.74 1072. 9 0.56 06 0.25 1077. 2 1.20 12 0.28 1091. 3 1.89 19 0.40 1101. 3 1.44 15 0.49 1112. 9 1.93 0.20 0.40 1126. 8 3.91 0.40 0.77 1142. 5 1.39 0.14 0.31 1155. 9 11.72 1.20 2.70 1171. 1 89 0.19 0.28 1178. 2 26 0.74 1.65 1194. 2 23 0.64 2.05 1202. 0 53 0.05 0.34 1207. 9 00 0.10 0.43 1215. 8 36 0.34 0.70 1226. 8 51 0.15 0.43 TJ 1231. 4 61 0.16 0.52 TJ 1237. 0 48 0.15 0.34 1245. 4 99 10 0.43 1258. 8 0.91 09 0.28 1274. 6 1.13 12 0.34 1287. 4 62 68 1.93 1295. 9 14 0.12 0.28 1303. 1 47 0.05 0.25 1309. 9 08 0.11 0.43 1325. 7 12 0. 83 1.04 1342. 0 50 0. 05 0.28 1351. 3 2.27 0.23 0.31 1367. 0 1.31 0.13 0.34 1370. 9 1.58 0 16 0.31 1386. 7 1.11 0 11 0.43 1409. 0 1.81 0 18 0.31 1431. 4 1.15 0 12 0.28 1444. 0 2.98 0.31 0.52 1456. 6 1.13 12 0.40 1475. .4 1.26 13 0.40 1492. .9 3.55 36 0.49 1500. .2 0.74 08 0.28 1507. .5 2. .33 24 0.31 1534. .8 0.56 06 0.25 1556. .8 4..03 41 0.52 1605. .3 2,.06 21 0.37 1626. .6 2..13 22 0.52 1642. .6 1..44 15 0.40 1665. .0 1.41 14 0.31 1687. .0 3.14 32 0.67 1726. .5 0.63 06 0.28 1749 .0 0.73 0.07 0.28 1780 .8 0.57 0.06 0.28 lo Intact Cell MALDI Meningo 5/M96255510 Data: a0009.9 3 Jul 97 15:03 Cal: test 25 Apr 97 11 25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 58, tDE @ 1000 (bin 56) Intact Cell MALDI Men ngo 10/M97251540 Data: a0019.19 3 Jui 97 15:28 Cal: test 25 Apr 9 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 65, tDE @ 100C (bin 56) Mass Urea %Totai Apex(mV) Flags 512.3 1.58 0.28 0.67 516.4 0.59 0.11 0.46 520.7 0.53 0.09 0.46 522.9 0.46 0.08 0.43 U 526.3 22.76 4.05 6.95 U 541.0 5.96 1.06 2.45 U 544.5 100.00 17.80 36.34 J 552.9 14.75 2.62 3.62 U 567.0 12.62 2.25 6.00 582.9 22.52 4.01 8.64 TJ 588.9 3.49 0.62 1.59 TJ 605.5 11.51 2.05 5.64 621.4 6.45 1.15 2.54 U 627.6 4.14 0.74 1.75 U 643.3 3.72 0.66 1.10 659.5 5.81 1.03 1.84-ϋ 667.5 41.96 7.47 15.07 U 675.4 8.21 1.46 3.40 U 690.7 18.02 3.21 5.82 706.9 11.96 2.13 2.70 U 716.3 15.87 2.82 5.88 U 732.3 32.81 5.84 8.82 U 737.4 11.63 2.07 4.38 U 744.1 2.32 0.41 0.80 U 751.4 15.92 2.83 8.95 U 753.7 35.50 6.32 8.43 U 770.7 14.92 2.66 3.86 U 784.9 7.64 1.36 2.27 U 789.4 4.21 0.75 1.72 U 800.4 3.48 0.62 1.01 812.5 0.96 0.17 0.37 831.4 1.35 0.24 0.37 TJ 833.9 0.51 0.09 0.43 U 856.2 2.59 0.46 0.98 882.7 1.75 0.31 0.40 902.4 1.10 0.20 0.43 944.4 0.90 0.16 0.64 959.0 0.51 0.09 0.37 1006.4 0.60 0.11 0.34 1018.5 4.81 0.86 1.84 1055.9 6.23 1.11 2.18 U 1063.0 0.79 0.14 0.55 U 1077.5 1.06 0.19 0.49 1093.8 1.40 0.25 0.64 1100.0 1.16 0.21 0.37 1115.8 1.54 0.27 0.55 1130.3 0.87 0.16 0.37 1152.1 0.70 0.12 0.40 U 1157.5 3.17 0.56 1.65 U 1180.8 1.25 0.22 0.58 U 1185.4 1.47 0.26 0.52 U 1197.1 2.56 0.45 0.77 1211.6 0.40 0.07 0.34 1239.0 0.65 0.11 0.43 1289.1 1.39 0.25 0.61 1306.5 0.91 0.16 0.40 1414.6 1.08 0.19 0.34 Intact Cell MALDI Memngo 10/M97251540 Da t a : a0019.19 3 Jul 97 15:28 Cal: test 25 Aor 97 11 .- . ratos Kompact MALDI 2 V5.2.1 • ♦ Linear B t Mass Urea %Total ' " High P ° Wer : 6 =' tDE @ "00 ( bin 56 ) ( A Intact Cell MALDI Memngo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97_ 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power : 65 , tDE @ 1000 (bin 56) ags ^ Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16 14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5 2.1: + Linear High Power: 65, Mass %Area %Total Apex(mV) Flags tDE @ 1000 (bin 56) 919.6 1.40 0.05 0.12 931.9 2.76 0.09 0.15 937.7 1.72 0.06 0.18 945.6 2 17 0.07 0.15 948.2 0 50 0.02 0.12 956.6 2, 13 0.07 0.18 994 0, 59 0.02 0.12 998 0.32 0.01 0.09 1004 0.50 0.02 0.12 1017 0.72 0.02 0.12 1041 0.72 0.02 0.12 1057 0.50 0.02 0.09 1088 2.71 0.09 0.28 1096 2.71 0.09 0.15 1116 1.40 0.05 0.18 1132 0.50 0.02 0.12 1137 0.32 0.01 0.09 1156 2.44 0.08 0.21 1160 0.54 0.02 0.12 1163 3.35 0.11 0.31 1199 0.27 0.01 0.09 1224 0.32 0 01 0.09 1248 0.50 0 02 0 .12 1266 0.68 0.02 0.12 1274 1.22 0.04 0.15 1286 0.27 0.01 0.09 1298 1.27 0.04 0 .15 1307 0 50 0.02 0 .12 1323 0 45 0.02 0 .09 1338 1, 45 0.05 0 .18 1362 1.18 0.04 0 .12 1370 0.81 0.03 0 .15 1379 0.50 0.02 0 .12 1405 2.53 0.09 0 .18 1428.9 77 0. .03 0 ,12 1584.9 86 0..03 0 ,12 1644 .9 50 0.02 0 ,12 1718.6 63 0.02 0 09 1785.9 50 0.02 0 09 1790.7 54 0.02 0 12 1998.1 45 0.02 0 09 2073.2 27 0.01 0, 09 2137.0 0.45 0.02 0. 09 2217.8 1.36 0.05 0, 12 2227.6 0.27 0.01 0. 09 2231.2 0.95 0.03 0, 12 2235.6 2.22 0.07 0, 21 2248.2 1.00 0.03 0. 12 2266.6 86 0.03 0. 15 2275 54 0.05 0. 12 2292 62 0.09 0. 15 2299 81 0.09 0. 18 2317 04 0.04 0. 15 2323 94 0.13 0. 15 2332 0.45 0.02 0. 09 2336 2.40 0.08 0. 25 2342 0.86 0.03 0. 12 b Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1; + Linear High Power: 65, tDE @ 100C (bin 56) a Intact Cell MALDI Memngo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56) l Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 15: 14 Cal: test 25 Apr 97-11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56) ags tl Intact Cell MALDI Memngo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: ■ * ■ Linear High Power: 65, tDE ® 1000 (bin 56) Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 65, tDE @ 1000 (bin 56 ) Mass %Area Total Apex(mV) Flags 500.6 45.92 4.17 6.65 U 516.8 100.00 9.07 8.95 U 525.9 24.35 2.21 3.22 U 533.3 21.39 1.94 2.85 540.3 0.36 0.03 0.12 545.0 58.34 5.29 6.89 554.9 1.93 0.17 0.31 556.3 0.34 0.03 0.18 558.1 1.71 0.15 0.52 U 560.1 1.98 0.18 0.64 U 561.6 0.15 0.01 0.12 567.0 7.03 0.64 1.32 578.1 0.57 0.05 0.18 583.1 5.83 0.53 0.95 589.1 1.83 0.17 0.37 593.7 0.54 0.05 0.12 594.6 0.30 0.03 0.12 602.7 0.20 0.02 0.12 605.7 5.65 0.51 1.04 613.0 0.51 0.05 0.18 615.6 0.17 0.02 0.09 621.9 3.31 0.30 0.46 627.5 3.58 0.33 0.58 633.5 0.17 0.02 0.09 635.9 0.46 0.04 0.15 645.7 5.39 0.49 0.70 650.5 0.20 0.02 0.12 654.6 1.01 0.09 0.18 659.7 13.80 1.25 2.14 667.8 10.16 0.92 1.47 675.9 29.64 2.69 4.04 687.7 0.10 0.01 0.09 691.9 27.61 2.50 3.43 700.2 0.69 0.06 0.18 707.4 7.19 0.65 0.83 U 716.8 12.19 1.11 1.35 U 732.6 -2S—5Q 2.31 2.30 U 738.2 11.85 1.07 1.62 TJ 745.2 1.56 0.14 0.34 754.0 33.43 3.03 2.48 766.0 0.10 0.01 0.09 770.7 8.64 0.78 0.77 786.0 5.65 0.51 0.89 790.0 0.10 0.01 0.09 794.8 1.54 0.14 0.31 800.1 2.47 0.22 0.37 813.0 1.05 0.10 0.28 817.0 0.05 0.00 0.09 824.9 0.51 0.05 0.18 830.1 0.36 0.03 0.18 833.6 0.56 0.05 0.18 838.5 0.85 0.08 0.15 855.3 3.52 0.32 0.37 867.5 3.13 0.28 0.25 871.5 0.39 0.04 0.15 873.7 0.19 0.02 0.12 880.4 0.51 0.05 0.15 Intact Cell MALDI Memngo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 ( b i n 56 ) Mass %Area Total Apex(mV) Flags 883.2 2.25 0.20 0.18 893.7 0.63 0.06 0.12 898.7 1.10 0.10 0.25 905.6 0.19 0.02 0.12 913.8 C.19 0.02 0.09 915.6 0.27 0.02 0.12 923.9 2.16 0.20 0.18 927.9 0.71 0.06 0.18 942.4 2.62 0.24 0.28 952.6 0.51 0.05 0.15 956.6 0.19 0.02 0.12 966.3 1.81 0.16 0.28 980.2 3.16 0.29 0.25 998.3 0.32 0.03 0.12 1001.6 1.94 0.18 0.28 1010.6 0.12 0.01 0.09 1014.2 0.29 0.03 0.15 1018.8 1.35 0.12 0.31 1040.3 0.10 0.01 0.09 1042.7 0.71 0.06 0.21 1051.9 0.19 0.02 0.12 1056.8 4.02 0.36 0.58 1063.9 0.27 0.02 0.12 1078.8 2.71 0.25 0.46 1085.9 0.29 0.03 0.09 1091.6 0.52 0.05 0.21 1094.7 0.98 0.09 0.15 1100.3 0.83 0.08 0.21 1117.0 2.05 0.19 0.18 1121.8 0.78 0.07 0.28 1128.4 0.17 0.02 0.09 1132.2 2.60 0.24 0.34 1146.9 0.49 0.04 0.12 1153.3 0.17 0.02 0.09 1158.8 2.71 0.25 0.43 1164.0 0.98 0.09 0.18 1170.1 0.12 0.01 0.09 1175.3 1.44 0.13 0.2& 1180.2 0.57 0.05 0.21 1189.3 1.37 0.12 0.25 1192.5 0.12 0.01 0.09 1204.7 0.47 0.04 0.12 1208.6 0.36 0.03 0.12 1210.6 0.12 0.01 0.09 1227.1 0.98 0.09 0.31 1234.7 0.34 0.03 0.12 1254.4 0.63 0.06 0.18 1259.1 0.79 0.07 0.18 1287.8 0.85 0.08 0.18 1292.9 1.35 0.12 0.28 1304.8 0.19 0.02 0.12 1315.1 0.19 0.02 0.12 1330.9 0.44 0.04 0.18 1339.9 0.56 0.05 0.21 1346.1 0.36 0.03 0.15 1375.4 0.39 0.04 0.15 1401.2 0.27 0.02 0.12 Ή . Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56 ) Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE ? 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 2447.5 0.85 08 0.18 2452.6 0.79 07 0.28 2460 3.31 30 0.25 2476 0. 10 01 0.09 2483 0.42 04 0.15 2490 0.51 05 0.18 2495 0.29 03 0.12 2499 0.42 04 0.15 2507.8 0.32 03 0.12 2518.7 1.91 0.17 0.28 2531.5 0.17 0.02 0.09 2539 0.51 0.05 0.18 2548 8.59 0.78 0.37 2582 0.59 0.05 15 2590 0, 83 0.08 .15 2599 1.50 14 31 2604 0.17 02 .09 2608 0.81 07 .21 2614.5 0, 44 04 ,15 2632.4 3.21 29 0.18 2645.5 5 17 47 46 2660.6 3 92 0.36 49 2673.3 0 10 0.01 09 2682.1 2 03 0. 18 28 2693 3 55 0.32 37 2703 0.10 0.01 09 2711 64 0.06 15 2717 20 0.11 28 2728.4 00 0.18 0.18 2736.3 86 0, 08 0.21 2741 .2 42 0.04 0.12 2750.7 20 0.20 0.28 2759.1 24 0.02 0.09 2766.6 64 0.06 0.15 2772.5 25 0.02 0.12 2777.0 62 0.15 0.25 2786.5 -CU3.fi. 0.03 0.12 2793.0 1.71 0.15 0.21 2805.0 2.23 0.20 0. 34 2814.5 2.59 0.23 0.18 2834.6 0.32 0.03 0.12 2839.2 0.37 03 0.15 2843.7 54 14 0.25 2851.8 25 02 0.12 2862.4 87 53 0.64 2876.6 ,00 0.09 0.25 2892.3 ,50 0.50 0.46 2911.2 .45 0.13 0.21 2917.3 ,10 0.01 0.09 2920.9 .51 0.05 0, 15 2928.6 .39 13 0.18 2937.8 .25 02 0.12 2945 .67 15 0 25 2954 .85 08 0 21 2963 .64 33 0 ,40 2977 .13 19 0.31 2990 2.98 27 0.28 ?3 Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56) Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56) Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE @ 1000 (bin 56) U Intact Cell MALDI Memngo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE ? 1000 ( b in 56)

Claims: CLAIMS 1. A method of characterising micro-organisms characterised by:- providing a database of matrix assisted laser desorption ionisation time of flight mass spectrometry (MALDI-TOF-MS) spectral data for a range of known micro-organisms. preparing a sample of unidentified micro-organisms and obtaining spectral data thereof by MALDI-TOF-MS and comparing using suitable comparison means the spectral data so obtained with database to identify a known micro-organism having the same or similar spectral data. 2. Apparatus for the screening of micro-organisms characterised in that the apparatus comprises :- spectrometric means for producing MALDI-TOF-MS spectral data of a sample organism database means containing MALDI-TOF-MS spectral data for a range of micro-organisms and comparison means for comparing the spectral data of the sample with that of the database to permit classification/identification of the sample. 3. Method or apparatus as claimed in claim 1 or claim 2 characterised in that the comparison means is a computer. 4. A method or apparatus as claimed in any preceding claim characterised in that the database means comprises spectral data recorded as a graphical representation of plot intensity against mass to charge ratio. 5. A method or apparatus as claimed in any preceding claim characterised in that the database means comprises spectral data recorded in graphic, numerical, or in electronic format in digital or analogue form and in that the comparison means is configured accordingly. 6. A method or apparatus as claimed in any preceding claim characterised in that the spectral data in the database is arranged in groups of data according to the genus of each micro-organism, with sub-divisions corresponding to each strain of micro-organism. 7. A method or apparatus as claimed in any preceding claim characterised in that the sample of unidentified organism is prepared either by taking cells from a culture and applying them to a sample plate comprising a matrix or by admixing the cells with the matrix prior to subjecting to MALDI-TOF-MS analysis in order to retain the cellular integrity of the sample. 8. A method or apparatus as claimed in any preceding claim characterised in that a sample matrix mixture is prepared and is bombarded with laser energy to create a gas phase ionic species which are then pulsed in to a flight conduit or tube for identification of both positive and/or negative ions. 9. A method or apparatus as claimed in any preceding claim characterised in that each species present is identified by their mass/charge ratio. 10. A method or apparatus as claimed in claim 9 characterised in that the mass/charge ratio of each spectral peak is determined from the centroid of the peak corresponding to the average molecular mass of the particular ion. 11. A method or apparatus as claimed in any preceding claim characterised in that the spectral data is derived from a plurality of laser shots of the sample in which the position and/or energy of the radiation impinging on the sample is varied between shots for the same sample. 12. A method or apparatus as claimed in any preceding claim characterised in that linear analysing is used to enhance sensitivity of data. 13. A database of micro-organisms incorporating spectral data obtained by MALDI-TOF-MS analysis of known microorganisms for use in the method or apparatus claimed in any preceding claim. 14. A database as claimed in claim 13 comprising at least one of the following organisms :- E.coli 8110 E.coli 10214 K12 lambda K.aerogenes 8172 C.freundii 9750 S.aureus Bl S.aureus B2 S.aureus methicillin resistant (MRSA) 01/270030 S.aureus (MRSA) 02/270238 *C.difficile NCTC 11204 *C.difficile NCTC 11206 *C . perfringens *C.tetani M.malmoense 0036c M.avium-intracellulare-scrofulaceum 0352c M. avium-intracellulare 0589c M.xenopi 0545c Nesseria species M96255506 Nesseria species M96255510 Nesseria species M97251540 Nesseria species M97251545 Nesseria species M97251682 ich the spectral data is as follows &l Intact Cell MALDI KRATOS DB Set E.coli 8110 Data: stdOOOl.l 24 Jul 97 16:26 Cal: -july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) 1232.8 100.00 10.99 35.48 S3 Intact Cell MALDI KRATOS DB Set E.coli 8110 Data: stdOOOl.l 24 Jul 97 16:26 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56] :uly97v3 24 Jul 97 11:05 » ar High Power: 60, tDE @ 1000 (bin 56) ss Intact Cell MALDI KRATOS DB Set E.coli 10214 K121am Data: std0003.3 24 Jul 97 16:28 Cal: july97v3 24- Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) i Intact Cell MALDI KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56 ) KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: :uly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56 ) 22 Intact Cell MALDI KRATOS DB Set K.aerogenes 8172 Data: std0013.13 24 Jul 97 16:39 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 2388.8 1.10 0.24 0.43 2402.2 0.67 0.15 0.43 Intact Cell MALDI KRATOS DB Set C.freundii 9750 Data: std0015.15 24 Jul 97 16:42 Cal: july97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE @ 1000 (bin 56) 30 Intact Cell MALDI KRATOS DB Set C.freundii 9750 Data: std0015.15 24 Jul 97 16:42 Cal: ]uly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 VS.2.1: + Linear High Power: 60, tDE § 1000 (bin 56) 3-f Intact Cell MALDI KRATOS DB Set C.freundii 9750 Data: std0015.15 24 Jul 97 16:42 Cal: 3 uly97v3 24 Jul 97 11:05 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60^ tDE S 1000 (bin 56 ) Ξ. aureus isolates Nutrient Agar Bl Data: aOOOl.l 17 Jul 97 17:18 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56 ) 53 S . aureus isolates Nutrient Agar Bl Data: aOOOl.l 17 Jul 97 17:18 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 70, tDE @ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 1388.1 0.91 0.19 0.46 TJ 1426.4 2.40 0.51 0.64 1458.0 3.16 0.68 0.61 1473.2 1.29 0.28 0.55 1518.6 3.02 0.64 0.43 1534.8 1.69 0.36 0.55 1554.9 1.75 0.37 0.52 U 1569.1 1.68 0.36 0.52 TJ 1827.3 1.01 0.22 0.55 1841.9 1.00 0.21 0.46 2000.7 2.13 0.45 0.43 2100.5 2.59 0.55 0.43 2387.0 1.59 0.34 0.46 2516.3 1.38 0.29 0.49 2814.5 3.14 0.67 0.46 5 S. aureus isolates Nutrient Agar B2 Data: a0003.3 17 Jul 97 17:22 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE & 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 500.8 74 0. 57 1.35 505.5 19 0. 14 0.61 512.3 28 1. 11 62 516.8 15 0. 62 29 526 17 1. 46 67 530 15.58 1. 87 57 543 45.57 5. 48 52 552 .29 0. 16 74 557 .15 0. 50 29 563 20.99 2. 52 4.50 569 81 0. 46 1.44 575 84 0. 58 1.16 581 27.92 3. 35 4.14 596 4.15 0. 50 0.77 603 11.88 1. 43 1.59 620 9.11 1. 09 1.93 622.6 4.52 0. 54 1.78 625 5.06 0. 61 1 62 635 4.64 0. 56 1 10 645 6.06 0. 73 1 10 650.8 3.37 0. 41 0 95 659.5 32.49 3. 90 5 82 666.8 21.15 2. 54 4 07 674.4 71 60 60 17 83 684 .2 5, 63 0. 68 1 44 690.4 100.00 12. .02 22 86 706.4 47.63 5. .72 11.64 715.3 .34 0. .04 0.21 722.6 .47 0. .78 1.23 731.3 .10 0. .13 0.49 741.6 1-40 0. .17 0.40 746.5 4.67 0. .56 0.74 757.4 .39 0. .05 0.21 764.1 .45 0. .17 0.37 772. 11.04 1. .33 78 790. 22.67 2. .72 76 807. 10.23 1. .23 99 u 811. 4.49 0. .54 16 TJ 819. 26 0 .03 21 827 .1 52 0 .90 23 u 833..9 85 0 .46 95 U 846..0 23.74 2 .85 98 86 .2 6.84 0 .82 1.90 U 869.2 88 0 .71 1.13 U 880.4 84 0 .58 1. 07 886.0 39 1 .13 2. 39 894.2 36 0 .28 0. 46 902.4 19.79 2 .38 3. 52 911.0 3.42 0 .41 1. 04 918.4 8.74 1 .05 1.56 937.1 4.28 0 .51 0.55 953.4 2.61 0 .31 0.55 963.7 55 0 .43 0.49 982.9 70 0 .56 0.46 1000.7 85 0 .46 0.52 1015.4 52 0 .30 0.61 1021.2 64 0 .44 0.70 as 5. aureus isolates Nutrient Agar B2 Data: a0003.3 17 Jul 97 17:22 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 70, tDE @ 1000 (bin 56) lags % S. aureus isolates Nutrient Agar B2 Data: a0003.3 17 Jul 97 17:22 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE <? 1000 (bin 56) 3 Intact Cell MALDI MRSA 01 /270030 Data: a 0001.1 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) Mass %Area %Total Apex( V) Flags 502. 0.35 0.07 0.12 517. 0.35 0.07 0.12 533. 0.29 0.06 0.12 536. 0.73 0.15 0.18 558. 0.62 13 0.21 565. 0.44 09 0.18 581. 23 05 0.12 582. 41 09 0.18 584. 35 0.07 0.12 596. 83 0.17 0.15 599. 15 0.03 0.09 601. 15 0.03 0. 09 622. 25 0.05 0. 12 630. 0.23 0.05 0. 12 633. 0.12 0.03 0. 09 637. 0. 12 0. 03 0. 09_ 639. 0.97 0.20 0. 31 655. 2.49 0.52 0. 28 659. 4.52 0.94 0. 83 662 0.15 0.03 0. 09 671. 5.70 1, 18 0. 86 675 35, 98 7.48 5. 02 687. 0, 89 0, 19 0.25 691. 84 81 17, 63 11.40 700. 0.15 0, 03 0.12 707. 100.00 20, 78 10.78 723. 30 72 6, 38 3.92 732 0 58 0, 12 0. 12 753. 62 0.13 0.12 763. 12 0, 23 0.18 766 06 0 01 0.09 767 12 0 03 0.09 774 77 0 16 0.18 777. 44 0 09 0.18 779. 21 0 04 0.09 787. 47 0 31 0.34 792 0.15 0 03 0.09 794 0.15 0 03 0.09 803 0.79 0 16 0.25 813 0.25 0 05 0.12 817. 0.12 0 03 0.09 819 0.15 0 03 0, 09 824 0.87 0 18 0.18 831. 0.21 0 04 0 12 834 0.54 0.11 0.18 846 0.23 05 0 12 848 0.33 07 0.12 849 0.91 19 0.28 855 0.17 03 12 861 0.89 19 18 865 1.62 34 28 871 0.37 0.08 15 876 0.46 0.09 12 877 0.23 0.05 12 881 3.83 0.80 0.46 887 2.61 0.54 0.58 897 5.16 1.07 0.77 Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) 55 Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 + Linear High Power: 70, Flags tDE @ 1000 (bin 56) .48 10 0.15 .23 05 12 .31 06. 12 .54 11 18 .12 0.03 09 ,23 05 12 .73 15 15 .21 04 0.09 ,31 06 0.12 0.23 0.05 0.09 oo Intact Cell MALDI MRSA 01/270030 Data: aOOOl.l 3 Jul 97 17:50 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: * ■ Linear High Power: 70, tDE & 1000 (bin 56) OΛ Intact Cell MALDI MRSA 02/270238 Data: a0002.3 3 Jul 97 17:53 Cal: test 25 Apr 97-11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 70, tDE @ 1000 (bin 56) <iθl* Intact Cell MALDI c. diff NCTC 11204 Data: diff0032.12 18 Jun 97 15:11 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 VS.2.1: - Linear High Power: 90, tDE @ 1000 (bin 56) Mass %Area %Total Apex( V) Flags 529.0 15. 84 541.9 3. .93 545.2 0. ,31 546.8 0. .45 550 0. .48 568 1. .46 585 0. .95 597 1.34 613.7 1.60 20.23 5.83 9.17 0.34 100.00 17.37 4.01 0. 83 1 .75 0.86 2.30 52.84 88.63 1 88 3.80 0 90 1.87 1 04 2.45 10 98 20.68 3 31 6.74 3 17 7.78 91 96 127.14 U 29 54 61 03 7 40 16 76 U 0 90 2 18 TJ 1 36 2 08 0 β2 .20 1 56 u 1 98 .49 3 62 1 19 .29 2 11 1 04 .26 1.96 8.76 .15 15.23 TJ .39 .10 1.59 .57 .39 2.88 U 0.72 .18 2.02 TJ Intact Cell MALDI c. diff NCTC 11206 Data: dιff0033.13 18 Jun 97 15:12 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1. - Linear High Power: 90, tDE @ 1000 (bin 56) ags Intact Ceil MALDI c. diff PERF Data: diff0037.17 18 Jun 97 15:19 Cal: test 25 Apr 97 11:31 Kratos Kompact MALDI 2 V5.2.1 : - Linear High Power: 95, tDE <? 1000 (bin 56) AoS t? 1000 (bin 56) - θ(> Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75/ tDE @ 3819 (bin 109) AoT- Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, tDE @ 3819 (bin 109) JoS Intact Cell MALDI Myco Cardiff l/0036c Data: CAR0002.2 30 May 97 17:06 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power : 75, tDE & 3819 (bin 109) Mass %Area %Total Apex(mV) Flags 2507.3 7.93 1.59 U 2522.0 8.49 1.84 U 2539.1 11.36 1.56 2579.8 7.17 1.29 2604.8 9.66 1.53 2620.8 5.57 1.19 2644.1 2.89 0.98 U 2657 11.59 2.02 U 2676 7.07 1.01 U 2710 25.00 3.86 U 2724 4.58 1.50 U 2769 9. 57 1.72 2789 4.88 1.04 2805 15.85 2.76 2864 10.04 1.47 2900 2.35 0.49 2948.5 15.82 2.30 3064 4 49 0.40 3111 2 87 61 _*7 4 00 61 U . ' . ' 7 3 42 55 U 32C7 2 22 58 3224 3 25 52 3250 0 98 25 3340 4.66 46 3378 0.74 28 7634.5 29.92 14 -tes Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16:27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE θ 1000 (bin 56) ■ HO Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16 27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 VS.2 1: + Linear High Power: 75, tDE @ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 1895.9 1 31 0.13 0.92 U 1902.9 1 12 0.11 0.86 U 1926.1 1 62 0.16 1.44 TJ 1931 2 54 0.25 2 . 05 u 1947 17 04 1.71 8 , 64 TJ 1961 7 84 0.79 4 . . 93 TJ 1974 100 ,00 10.04 52 , . 36 TJ 1989 59 .98 02 32 . 94 TJ 2002 46 .84 70 26 . 07 U 2017 62.90 32 35 .97 2032.1 20.67 08 13 . 27 2044.5 16 82 69 10 .94 2059.9 7 32 0.74 5 . 33 2073.2 73 0.17 2088.3 39 14 2109 66 27 2121 56 36 2152 18 52 2164 2 95 70 2175 ,7 13 42 2192.0 62 16 2216.9 62 46 2234.3 1.47 15 2243.3 2.61 26 2252.2 4.99 50 2266.6 1.88 19 2278.3 2.03 20 2291.0 1.43 14 2299.6 43 14 2307.3 22 32 2325.1 07 0.11 2339.7 68 0.27 2354.4 79 0.38 2370.0 91 0.59 2388.9 4.53 0.45 2416.7 2.13 0.21 2432.1 6.65 0.67 2464.4 2 95 0.30 2479.4 6 16 0.62 2504.0 4 20 42 2527.2 2 68 27 2538.2 5 39 54 2568.3 4 26 43 2592.8 4 20 42 2618.8 2 39 24 2633 ,9 2 35 0.24 2661 , i 7 83 0.79 2704 .7 66 01 6.63 2 2751.2 1.94 0.20 2762.6 5.71 0.57 2795 .5 72 0.17 2815.0 04 0.20 2825.6 13 0.21 2849.3 40 0.54 2869.5 28 0.33 2894.9 1.56 0.16 2919.9 2.12 0.21 H Intact Cell MALDI Myco Cardiff 31/0352c Data: CAR0082.2 3 Jun 97 16:27 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, tDE @ 1000 ( bin 56 ) Mass %Area %Total Apex( V) Flags 2928.6 1.05 0.11 0.86 U 2943.4 4.14 0.42 1.65 TJ 2967.1 3.05 0.31 1.23 U 2981.5 1.00 0.10 0.64 U 2995.0 1.86 0.19 0.89 U 3018.3 2.83 0.28 1.35 U 3032.9 0.86 0.09 0.64 U 3054.8 4.50 0.45 0.92 U 3092.0 2.53 0.25 0.86 TJ 3108.9 3.68 0.37 1.78 U 3125.2 1.38 0.14 0.61 TJ 3148.5 4.60 0.46 1.65 TJ 3167.7 1.62 0.16 0.67 U 3200.8 2.17 0.22 0.61 TJ 3237.2 4.58 0.46 1.26 U 3276.6 2.28 0.23 0.95 U 3332.1 3.02 0.30 1.07 U 3362.7 5.16 0.52 0.98 TJ 3422.2 16.59 1.67 5.88 TJ 3449.4 1.63 0.16 0.52 U 3490.1 3.20 0.32 0.74 3591.4 1.83 0.18 0.52 U 3642.0 3.64 0.37 0.83 U 3709.7 3.62 0.36 0.58 U 3772.7 2.57 0.26 0.64 3811.7 2.13 0.21 0.58 TJ 3871.5 1.39 0.14 0.52 U 3896.9 3.04 0.31 1.35 U 3914.0 2.60 0.26 1.07 TJ 3933.0 2.07 0.21 0.83 TJ 3974.6 1.33 0.13 0.58 U 3987.1 1.75 0.18 0.61 U 4859.8 3.58 0.36 0.77 Intact Cell MALDI Myco Cardiff 33/0589c Data: CAR0086.6 3 Jun 97 16:37 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) Intact Cell MALDI Myco Cardiff 33/0589c Data: CAR0086.6 3 Jun 97 16:37 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 : + Linear High Power: 75, tDE & 1000 (bin 56) Ηl\ Intact Cell MALDI Myco Cardiff 33/0589c Data: CAR0086.6 3 Jun 97 16:37 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 ( b in 56) -ΉS Intact Cell MALDI Myco Cardiff 63 /0545c Data: CAR0146.6 4 Jun 97 14:12 Cal: test 25 A pr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, t D E β 1000 ( b in 56 ) Intact Cell MALDI Myco Cardiff 63/0545c Data: CAR0146.6 4 Jun 97 14:12 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 75, tDE @ 1000 (bin 56) tMt „? 5 Apr 97 11:25 Hl9h PθWer: 75 ' tDE • "00 ( bin 56; ■ 8 Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 + Linear High Power: 60, tDE θ 1000 (bin 56) -MS Intact Cell KALDI Mβningo 1/M96255506 9 1000 (bin 56) Λ2o Intact Cell MALDI Meningo 1/M96255506 tDE 9 1000 (bin 56) AIΛ Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 VS.2.1: + Linear High Power: 60, tDE 9 1000 (bin 56) Flags Zl Intact Cell MALDI Meningo 1/M96255506 Data: a0002.2 3 Jul 97 14:39 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 60, tDE θ 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 5631.3 1.48 0.19 0.18 5683.2 2.46 0.32 0.18 5701.8 1.72 0.22 0.15 5721.8 0.43 0.06 0.09 6013.8 6.28 0.81 0.37 6033.6 2.55 0.33 0.21 6069.6 11.02 1.42 0.55 6091.0 3.75 0.48 0.25 6151.0 1.02 0.13 0.12 23 Intact Cell MALDI Meningo 5/M96255510 Data : a0009.9 3 Jul 97 Kratos Kompact MALDI 2 tDE 9 1000 (bin 56) Mass %Area %Total 500. 16.10 1.65 509. 0.96 0.10 511. 1.68 0.17 516. 100.00 10.24 520. 6. 50 0.67 525. 16.29 1.67 532. 86.19 8.82 543. 66.76 6.84 551. 4.52 0.46 553. 5.57 0.57 558. 1.44 0.15 563. 3.49 0.36 566. 10.84 1.11 575. 0.84 0.09 582. 17.18 1.76 587. 0.73 0.07 598. 0.59 0.06 604. 10.71 10 620. 14.90 53 633. 1.87 19 636. 0.83 09 644. 5.86 60 652. 0.33 0.03 659. 13.30 1.36 666. 22 ,73 2.33 674. 24.07 2.46 690. 69.23 7.09 706. 27.36 2.80 715. 20.15 06 731. 68.65 03 746. 8.86 91 750. 24.51 51 753. 24.65 52 6.16 757. 5.85 60 2.42 762. 19.36 1.98 3.83 769. 15.50 1.59 4.78 773. 17.49 1.79 3.71 784. 20.78 2.13 3.03 801. 14.75 1.51 2.51 810. 3.13 0.32 0.64 825 1.56 0.16 0.58 828 4.13 0.42 0.89 836 0.75 0.08 0.31 840 0.67 0.07 0.28 855. 3.76 0.38 0.61 865 2.61 0.27 0.43 880. 2.94 0.30 0.64 897 3.75 0.38 0.86 902 58 0.26 0.83 907 61 0.06 0.31 913 77 0.18 0.52 920. 19 0.22 0.61 930 98 0.20 0.37 936 0.52 0.05 0.28 942 6.94 0.71 1.41 956 5.84 0.60 1.59 967 1.42 0.15 0.34 ^2c, Intact Cell MALDI Meningo 5/M96255510 Data: a0009.9 3 Jul 97 15:03 Cal: test 25 Apr 97 11: 25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 58, tDE 9 1000 (bin 56) 25 test 25 Apr 97 11:25 inear High Power: 58, tDE 9 1000 (bin 56) ags ΛZi Intact Cell MALDI Meningo 10/M97251540 Data: a0019.19 3 Jul 97 15:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) Intact Cell MALDI Meningo 10/M97251540 Data: a0019.19 3 Jul 97 15:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56 ) Mass %Area %Total Apex(mV) Flags 1446.5 0.56 0.10 0.34 1471.8 0.92 0.16 0.43 1560.5 1.16 0.21 0.40 1607.2 1.87 0.33 0.98 1635.4 0.80 0.14 0.37 1644.9 1.55 0.28 0.67 1689.7 1.16 0.21 0.46 2692.9 1.24 0.22 0.49 2862.9 1.39 0.25 0.77 3313.0 1.07 0.19 0.40 5145.4 1.08 0.19 0.37 6424.7 2.65 0.47 0.64 6520.6 8.67 1.54 1.93 6619.5 46.42 8.26 8.58 D 6657.2 6.34 1.13 1.47 ϋ 6686.6 0.85 0.15 0.46 U 6695.9 1.04 0.18 0.43 ϋ -fU? Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) ags -fZS Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE β 1000 (bin 56) -ι3o Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) - 3-f Intact Cell MALDI Meningo 15/M97251545 Data : a0029 .9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE i 1000 (bin 56) Mass %Area %Total Apex(mV) Flags 3194.9 3.44 0.12 0.31 3204.5 6. 83 0.23 0.61 3219.5 32 .26 1.09 0.61 U 3244.2 26 .92 0.91 0.61 U 3283.7 18 .55 0.63 0.58 3308.1 11.49 0.39 0.46 3331.0 16.15 0.55 0.40 3339.7 5.52 0.19 0.49 U 3349.0 13 .76 0.46 0. .77 ϋ 3367.1 14.25 0.48 0. .55 ϋ 3379.2 11.18 0.38 0. .58 U 3401.2 27.51 0.93 0.58 U 3421.7 14.93 0.50 0. .61 U 3435.5 9 .91 0.33 0.49 U 3442.2 8.28 0.28 0.55 U 3450.0 10.45 0.35 0.52 3465.0 9.91 0.33 0.49 3473.3 10.63 0.36 0.64 3490.1 16.43 0.55 0.67 3511.3 26.65 0.90 0.74 3529.9 1.86 0.06 0.18 3554.6 32.08 1.08 0.74 3569.3 16.15 0.55 0.49 3591.9 13.03 0.44 0.67 3605.6 14.39 0.49 0.43 3628.9 22.49 0.76 0.80 3647.7 15.97 0.S4 0.70 U 3666.0 34.07 1.15 0.74 U 3683.2 10.90 37 0.58 ϋ 3694.7 1.09 04 0.12 3716.6 25.43 86 04 ϋ 3731.0 12. .31 42 52 U 3750.1 17..69 60 74 3766.3 13..98 0.47 49 U 3779.1 18..51 0.63 0.95 U 3794.8 25..66 0.87 0.92 U 3818.8 9.77 0.33 0.40 3870.9 - ^ 03 1.79 0.61 U 3881.0 6.56 0.22 49 IT 3893.3 12.08 0.41 61 U 3908.1 8.60 0.29 40 3927.6 23.67 0.80 52 3948.4 1.95 0.07 0.15 3962.7 8.96 0.30 0.52 U 3983.0 20.41 0.69 0.43 U 4006.3 15.38 0.52 0.40 U 4046.5 30.14 1.02 0.70 U 4073.0 10.09 0.34 0.64 U 4094.2 16.43 0.55 0.55 U 4109.3 16.38 0.55 0.55 U 4133.0 12.49 0.42 37 4156.7 10.27 0.35 46 U 4171.4 17.24 0.58 64 U 4185.4 11.40 0.39 64 u 4205.7 12.22 0.41 0.58 U 4220.4 8.69 0.29 0.40 U 4237.0 18.24 0.62 0.52 u IZ Intact Cell MALDI Meningo 15/M97251545 Data: a0029.9 3 Jul 97 16:14 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE β 1000 (bin 56) 33 : test 25 Apr 97 11: 25 Linear High Power: 65, tDE 9 1000 (bin 56] Flags l | Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) -MS Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) Flags 3 Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE β 1000 (bin 56) lags -t37- Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1 + Linear High Power: 65, tDE 9 1000 (bin 56) Flags 438 test 25 Apr 97 11:25 Linear High P ower: 65, lags tDE 9 1000 (bin 56) -133 Intact Cell MALDI Meningo 20/M97251682 Data: a0039.19 3 Jul 97 16:28 Cal: test 25 Apr 97 11:25 Kratos Kompact MALDI 2 V5.2.1: + Linear High Power: 65, tDE 9 1000 (bin 56) ags tDE & 100 ° (bιn 56 )

Inventors: CLAYDON MARTIN ARTHUR, DAVEY SIMON NICHOLAS, EDWARDS JONES VALERIE, GORDON DEREK BOYD, CLAYDON, MARTIN, ARTHUR, DAVEY, SIMON, NICHOLAS, EDWARDS JONES, VALERIE, GORDON, DEREK, BOYD
Applicants: CLAYDON MARTIN ARTHUR, DAVEY SIMON NICHOLAS, EDWARDS JONES VALERIE, GORDON DEREK BOYD, UNIV MANCHESTER METROPOLITAN, CLAYDON, MARTIN, ARTHUR, DAVEY, SIMON, NICHOLAS, EDWARDS JONES, VALERIE, GORDON, DEREK, BOYD, THE MANCHESTER METROPOLITAN UNIVERSITY
Application year: 1997
Main classification: H01J49
IPC codes: C12M1, C12N1, C12Q1, H01J49