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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).

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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

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#mAb Namesource SpecificityLinear Epitope Mapped N12-i9.1Human IgG1Neg CtrlNegative N12-i9.1Human IgG1Neg CtrlNegative 2F5Human IgG12F5 epitope 1N5-i5Human IgG1 gp120Negative 2N12-i3Human IgG1gp120Negative 3N26-i1Human IgG1gp120Negative 4L9-i1Human IgG1gp120gp41 5L9-i2Human IgG1gp120Negative 6JR48.1Human IgG1gp120C1 7JR49Human IgG1gp120C5 8A32Human IgG1gp120Negative 9C11Human IgG1gp120Negative 10N60-i3Human IgG1gp120Negative 11JR4Rhesus IgG1gp120Negative 12JR20Rhesus IgG1gp120Negative 13JR52Rhesus IgG1gp120C5 14N12-i15Human IgG1gp120C4 15N60-B1.1Human IgG1gp120Negative 16N5-U1Human IgG1gp41Negative 17N5-U2Human IgG1gp41Negative 18N5-U3Human IgG1gp41Negative 19N10-U1Human IgG1gp41gp41 ID, Clade C C1 & MPER 20N10-U2Human IgG1gp41gp41 CBD-1 21M785-U1Rhesus IgG1gp41gp41 ID 22M785-U2Rhesus IgG1gp41gp41 CBD-1 23M785-U3Rhesus IgG1gp41Negative 24M785-U4Rhesus IgG1gp41 gp41 CBD-1 S UMMARY OF R ESULTS

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N12-i9.1 Neg Ctrl

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N5-O1 Neg ctrl

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2F5 Pos Ctrl

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N5-i5

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N12-i3

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N26-i1

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L9-i1

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L9-i1 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT2049, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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L9-i2

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JR48.1

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JR48.1 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT2049, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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JR49

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JR49 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT1948, PMT540; SXS Background subtraction: 5-fold nonspecific peptide binding.

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A32

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C11

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N60-i3

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JR4

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JR20

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JR52

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JR52 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT1948, PMT580; SXS Background subtraction: 5-fold nonspecific peptide binding.

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N12-i15

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N12-i15 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT1948, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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N60-B1.1

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N5-U1

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N5-U2

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N5-U3

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N10-U1

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N10-U1 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot # JPT2049, PMT540; SXS 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.

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N10-U2

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N10-U2 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot# JPT1948, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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M785-U1

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M785-U1 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot# JPT1948, PMT580; SXS Background subtraction: 5-fold nonspecific peptide binding.

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M785-U2

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M785-U2 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot# JPT2049, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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M785-U3

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M785-U4

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M785-U4 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF µg/ml, Lot# JPT2049, PMT600; SXS Background subtraction: 5-fold nonspecific peptide binding.

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