40
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 Shaunna Shen/Georgia Tomaras, CAVIMC 24 mAb + 2 negative control tested: 20 µg/ml (default), 5, 10, or 40 µg/ml (for

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Page 1: 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

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).

Page 2: 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

• 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.

CAVIMC 040 mAb Linear Epitope Mapping

Page 3: 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

# mAb Name source Specificity Linear Epitope MappedN12-i9.1 Human IgG1 Neg Ctrl NegativeN12-i9.1 Human IgG1 Neg Ctrl Negative2F5 Human IgG1 2F5 epitope 2F5 epitope

1 N5-i5 Human IgG1   gp120 Negative2 N12-i3 Human IgG1 gp120 Negative3 N26-i1 Human IgG1 gp120 Negative4 L9-i1 Human IgG1 gp120 gp415 L9-i2 Human IgG1 gp120 Negative6 JR48.1 Human IgG1 gp120 C17 JR49 Human IgG1 gp120 C58 A32 Human IgG1 gp120 Negative9 C11 Human IgG1 gp120 Negative

10 N60-i3 Human IgG1 gp120 Negative11 JR4 Rhesus IgG1 gp120 Negative12 JR20 Rhesus IgG1 gp120 Negative13 JR52 Rhesus IgG1 gp120 C514 N12-i15 Human IgG1 gp120 C415 N60-B1.1 Human IgG1 gp120 Negative16 N5-U1 Human IgG1 gp41 Negative17 N5-U2 Human IgG1 gp41 Negative18 N5-U3 Human IgG1 gp41 Negative19 N10-U1 Human IgG1 gp41 gp41 ID, Clade C C1 & MPER20 N10-U2 Human IgG1 gp41 gp41 CBD-121 M785-U1 Rhesus IgG1 gp41 gp41 ID22 M785-U2 Rhesus IgG1 gp41 gp41 CBD-123 M785-U3 Rhesus IgG1 gp41 Negative24 M785-U4 Rhesus IgG1 gp41 gp41 CBD-1

SUMMARY OF

RESULTS

Page 4: 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

N12-i9.1 Neg Ctrl

N12-i9.1 (Neg Ctrl) 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

N12-i9.1 (Neg Ctrl) 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 5: 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

N5-O1 Neg ctrl

N5-O1 (Neg Ctrl) 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N5-O1 (Neg Ctrl) 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 6: 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

2F5 Pos Ctrl

2F5 10g/ml_gp41 IgG_PMT540_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

2F5 10g/ml_gp120 IgG_PMT540_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 7: 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

N5-i5

N5-i5 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

N5-i5 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 8: 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

N12-i3

N12-i3 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

N12-i3 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 9: 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

N26-i1N26-i1 10g/ml_gp120 IgG_PMT600_24Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N26-i1 10g/ml_gp41 IgG_PMT600_24Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPERImmunodominant

HR2

Kennedy Sequence LLP2

Page 10: 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

L9-i1L9-i1 10g/ml_gp120 IgG_PMT600_24Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

na

l In

ten

sit

y

0

2000

4000

6000

8000

10000

12000

14000Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

L9-i1 10g/ml_gp41 IgG_PMT600_24Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

na

l In

ten

sity

0

2000

4000

6000

8000

10000

12000

14000 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPERImmunodominant

HR2

Kennedy Sequence LLP2

Page 11: 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

L9-i1 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

200 295 0 0 0 0 0 0

201 0 1690 276 194 0 0 2138

202 0 0 148 0 0 0 0

203 0 0 0 0 62 0 0

214 1005 0 519 0 0 0 0

10 µg/ml, Lot # JPT2049, PMT600; SXS 23-11

Background subtraction: 5-fold nonspecific peptide binding.

Page 12: 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

L9-i2

L9-i2 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

L9-i2 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 13: 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

JR48.1JR48.1 20g/ml_gp120 IgG_PMT540_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

1.0e+5

2.0e+5

3.0e+5

4.0e+5

5.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5C5

C1

JR48.1 20g/ml_gp41 IgG_PMT540_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

500

1000

1500

2000

2500

3000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 14: 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

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 68 0 57

10 15016 6631 1432 4698 15016 12693 6680

11 23081 17655 7062 17655 23081 20499 4038

12 63124 46498 44927 46498 46498 49918 59875

13 566 689 469 689 689 787 0

10 µg/ml, Lot # JPT2049, PMT600; SXS 23-11

Background subtraction: 5-fold nonspecific peptide binding.

Page 15: 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

JR49JR49 20g/ml_gp120 IgG_PMT540_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

JR49 20g/ml_gp41 IgG_PMT540_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 16: 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

JR49 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

157 126 55068 0 22140 55068 259 0

158 163 44526 0 39524 44526 517 0

20 µg/ml, Lot # JPT1948, PMT540; SXS 23-06

Background subtraction: 5-fold nonspecific peptide binding.

Page 17: 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

A32

A32 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

A32 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 18: 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

C11

C11 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

C11 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 19: 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

N60-i3

N60-i3 20g/ml_gp41 IgG_PMT600_07Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

na

l In

ten

sity

0

2000

4000

6000

8000

10000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

N60-i3 20g/ml_gp120 IgG_PMT600_07Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

500

1000

1500

2000

2500

3000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 20: 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

JR4

JR4 (mk) 20g/ml_gp41 IgG_PMT600_27Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

JR4 (mk) 20g/ml_gp120 IgG_PMT600_27Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 21: 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

JR20JR20 (mk) 20g/ml_gp120 IgG_PMT600_27Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

JR20 (mk) 20g/ml_gp41 IgG_PMT600_27Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 22: 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

JR52JR52(mk) 20g/ml_gp120 IgG_PMT580_27Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

JR52(mk) 20g/ml_gp41 IgG_PMT580_27Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 23: 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

JR52 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

159 0 2688 0 0 0 0 0

160 13302 44634 13770 10031 12870 13357 11602

161 673 26587 482 288 482 673 115

162 0 12116 0 0 0 0 0

20 µg/ml, Lot # JPT1948, PMT580; SXS 23-09

Background subtraction: 5-fold nonspecific peptide binding.

Page 24: 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

N12-i15N12-i15 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

na

l In

ten

sit

y

0

2000

4000

6000

8000

10000

12000

14000 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N12-i15 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

na

l In

ten

sity

0

2000

4000

6000

8000

10000

12000

14000 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 25: 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

N12-i15 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

138 0 0 0 7099 0 0 0

20 µg/ml, Lot # JPT1948, PMT600; SXS 23-10

Background subtraction: 5-fold nonspecific peptide binding.

Page 26: 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

N60-B1.1

N60-B1.1 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

N60-B1.1 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 27: 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

N5-U1N5-U1 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N5-U1 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 28: 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

N5-U2N5-U2 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N5-U2 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 29: 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

N5-U3N5-U3 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N5-U3 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

1000

2000

3000

4000

5000

6000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 30: 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

N10-U1

N10-U1 5g/ml_gp41 IgG_PMT540_24Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

1.0e+5

2.0e+5

3.0e+5

4.0e+5

5.0e+5 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPERImmunodominant

HR2

Kennedy Sequence LLP2

N10-U1 5g/ml_gp120 IgG_PMT540_24Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

1.0e+5

2.0e+5

3.0e+5

4.0e+5

5.0e+5 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 31: 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

N10-U1 Signal Intensity ValuesPeptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

33 0 0 53415 0 0 0 0

189 58960 58960 58978 58960 58960 59156 59086190 59070 59070 59070 59070 59070 53422 59159194 0 0 0 13065 0 0 0195 5306 774 0 26403 5306 0 12130196 0 54161 47183 51288 0 44181 35031199 0 0 0 43418 0 0 0

214 0 0 40230 0 0 0 0

5 µg/ml, Lot # JPT2049, PMT540; SXS 23-11

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.

Background subtraction: 5-fold nonspecific peptide binding.

Page 32: 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

N10-U2N10-U2 20 g/ml_gp120 IgG_PMT600_16Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

2.0e+4

4.0e+4

6.0e+4

8.0e+4

1.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

N10-U2 20 g/ml_gp41 IgG_PMT600_16Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

2.0e+4

4.0e+4

6.0e+4

8.0e+4

1.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

CBD-1

Page 33: 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

N10-U2 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

198 0 0 0 0 0 0 0

199 0 843 0 8488 0 0 8634

200 0 21108 0 0 3140 0 2347

201 0 13024 13695 21420 9405 0 3795

202 0 0 5522 6134 7654 0 0

203 0 0 0 0 0 0 0

20 µg/ml, Lot# JPT1948, PMT600; SXS 23-10

Background subtraction: 5-fold nonspecific peptide binding.

Page 34: 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

M785-U1

M785-U1(mk) 20g/ml_gp41 IgG_PMT580_27Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

3.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

M785-U1(mk) 20g/ml_gp120 IgG_PMT580_27Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

3.0e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 35: 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

M785-U1 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

192 38178 38178 38178 0 38178 9730 38178193 16455 13054 13054 4931 16455 538 12801

194 22182 20079 21843 4928 22182 868 18114

20 µg/ml, Lot# JPT1948, PMT580; SXS 23-09

Background subtraction: 5-fold nonspecific peptide binding.

Page 36: 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

M785-U2

M785-U2(mk) 40g/ml_gp41 IgG_PMT600_24Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

na

l In

ten

sity

0

2000

4000

6000

8000

10000

12000

14000 Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPERImmunodominant

HR2

Kennedy Sequence LLP2CBD-1

M785-U2(mk) 40g/ml_gp120 IgG_PMT600_24Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

na

l In

ten

sit

y

0

2000

4000

6000

8000

10000

12000

14000Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

Page 37: 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

M785-U2 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

201 0 1285 0 2233 2403 0 0

40 µg/ml, Lot# JPT2049, PMT600; SXS 23-11

Background subtraction: 5-fold nonspecific peptide binding.

Page 38: 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

M785-U3M785-U3 (mk) 20g/ml_gp120 IgG_PMT600_27Mar2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sit

y

0

500

1000

1500

2000

2500

3000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

M785-U3 (mk) 20g/ml_gp41 IgG_PMT600_27Mar2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0

500

1000

1500

2000

2500

3000

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPER

Immunodominant

HR2

Kennedy Sequence

LLP2

Page 39: 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

M785-U4M785-U4(mk) 40g/ml_gp120 IgG_PMT580_24Apr2012

gp120 peptide number0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

V3

V2V1

V4 V5

C5

C1

M785-U4(mk) 40g/ml_gp41 IgG_PMT580_24Apr2012

gp41 peptide number

165 170 175 180 185 190 195 200 205 210 215 220 225 230 235 240 245 250 255 260 265 270 275

Sig

nal

In

ten

sity

0.0

5.0e+4

1.0e+5

1.5e+5

2.0e+5

2.5e+5

Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2 MPERImmunodominant

HR2

Kennedy Sequence LLP2

CBD-1

Page 40: 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

M785-U4 Signal Intensity Values

Peptide # Clade A Clade B Clade C Clade D Group M CRF 1 CRF 2

199 0 5598 0 0 1016 0 3438

200 2014 13458 2344 0 14384 2819 16795

201 1174 33507 27116 35009 33027 0 35006202 2552 7963 20051 23515 21665 0 2552

203 0 8507 0 8507 18555 0 0

40 µg/ml, Lot# JPT2049, PMT600; SXS 23-11

Background subtraction: 5-fold nonspecific peptide binding.