CAVIMC 040 mAb Linear Epitope Mapping

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Presentation transcript:

CAVIMC 040 mAb Linear Epitope Mapping Shaunna Shen/Georgia Tomaras, CAVIMC 24 mAb + 2 negative control tested: 20 µg/ml (default), 5, 10, or 40 µg/ml (for repeats/confirmation). Peptide sequences on slide: 15-mers overlapping by 12, covering full length consensus Env gp160 of HIV clade A, B, C, D, group M, and CRF 1 & 2. Slide Lots used: JPT 1948 (default) and JPT 2049 (for repeats/confirmation).

CAVIMC 040 mAb Linear Epitope Mapping X-axis: Peptide#, ordered from N’ to C’ on gp160. Y-axis: Signal intensity, as determined by image analysis of the chip scans. Colors of bars represent different clades/group/CRFs of HIV Env. Background subtraction: 5-fold the signal intensity for nonspecific peptide binding. Results are summarized in slide 3 as a table. Graphs of data for each mAb follow (Note: In the graphs, the y-axis scales differ based on response magnitude). Binding intensity values per clade for each mAb with positivity calls are shown in a table after the graph.

Summary of Results # mAb Name source Specificity Linear Epitope Mapped Specificity Linear Epitope Mapped N12-i9.1 Human IgG1 Neg Ctrl Negative 2F5 2F5 epitope 1 N5-i5 gp120 2 N12-i3 3 N26-i1 4 L9-i1 gp41 5 L9-i2 6 JR48.1 C1 7 JR49 C5 8 A32 9 C11 10 N60-i3 11 JR4 Rhesus IgG1 12 JR20 13 JR52 14 N12-i15 C4 15 N60-B1.1 16 N5-U1 17 N5-U2 18 N5-U3 19 N10-U1 gp41 ID, Clade C C1 & MPER 20 N10-U2 gp41 CBD-1 21 M785-U1 gp41 ID 22 M785-U2 23 M785-U3 24 M785-U4

N12-i9.1 Neg Ctrl

N5-O1 Neg ctrl

2F5 Pos Ctrl

N5-i5

N12-i3

N26-i1

L9-i1

L9-i1 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 200 295 201 1690 276 194 2138 202 148 203 62 214 1005 519 Background subtraction: 5-fold nonspecific peptide binding. 10 µg/ml, Lot # JPT2049, PMT600; SXS 23-11

L9-i2

JR48.1

JR48.1 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 9 68 528 130 346 57 10 15016 6631 1432 4698 12693 6680 11 23081 17655 7062 20499 4038 12 63124 46498 44927 49918 59875 13 566 689 469 787 Background subtraction: 5-fold nonspecific peptide binding. 10 µg/ml, Lot # JPT2049, PMT600; SXS 23-11

JR49

JR49 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 157 126 55068 22140 259 158 163 44526 39524 517 Background subtraction: 5-fold nonspecific peptide binding. 20 µg/ml, Lot # JPT1948, PMT540; SXS 23-06

A32

C11

N60-i3

JR4

JR20

JR52

JR52 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 159 2688 160 13302 44634 13770 10031 12870 13357 11602 161 673 26587 482 288 115 162 12116 Background subtraction: 5-fold nonspecific peptide binding. 20 µg/ml, Lot # JPT1948, PMT580; SXS 23-09

N12-i15

N12-i15 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 138 7099 Background subtraction: 5-fold nonspecific peptide binding. 20 µg/ml, Lot # JPT1948, PMT600; SXS 23-10

N60-B1.1

N5-U1

N5-U2

N5-U3

N10-U1

N10-U1 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 33 53415 189 58960 58978 59156 59086 190 59070 53422 59159 194 13065 195 5306 774 26403 12130 196 54161 47183 51288 44181 35031 199 43418 214 40230 Background subtraction: 5-fold nonspecific peptide binding. Note: “Breakage” of the gp41 ID epitope at peptide #192 and 193 is a known slide quality issue. The epitope is very likely a continuous one. 5 µg/ml, Lot # JPT2049, PMT540; SXS 23-11

N10-U2

N10-U2 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 198 199 843 8488 8634 200 21108 3140 2347 201 13024 13695 21420 9405 3795 202 5522 6134 7654 203 Background subtraction: 5-fold nonspecific peptide binding. 20 µg/ml, Lot# JPT1948, PMT600; SXS 23-10

M785-U1

M785-U1 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 192 38178 9730 193 16455 13054 4931 538 12801 194 22182 20079 21843 4928 868 18114 Background subtraction: 5-fold nonspecific peptide binding. 20 µg/ml, Lot# JPT1948, PMT580; SXS 23-09

M785-U2

M785-U2 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 201 1285 2233 2403 Background subtraction: 5-fold nonspecific peptide binding. 40 µg/ml, Lot# JPT2049, PMT600; SXS 23-11

M785-U3

M785-U4

M785-U4 Signal Intensity Values Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 199 5598 1016 3438 200 2014 13458 2344 14384 2819 16795 201 1174 33507 27116 35009 33027 35006 202 2552 7963 20051 23515 21665 203 8507 18555 Background subtraction: 5-fold nonspecific peptide binding. 40 µg/ml, Lot# JPT2049, PMT600; SXS 23-11