Presentation on theme: "CAVIMC 040 mAb Linear Epitope Mapping"— Presentation transcript:
1CAVIMC 040 mAb Linear Epitope Mapping Shaunna Shen/Georgia Tomaras, CAVIMC24 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).
2CAVIMC 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.
3Summary of Results # mAb Name source Specificity Linear Epitope Mapped SpecificityLinear Epitope MappedN12-i9.1Human IgG1Neg CtrlNegative2F52F5 epitope1N5-i5gp1202N12-i33N26-i14L9-i1gp415L9-i26JR48.1C17JR49C58A329C1110N60-i311JR4Rhesus IgG112JR2013JR5214N12-i15C415N60-B1.116N5-U117N5-U218N5-U319N10-U1gp41 ID, Clade C C1 & MPER20N10-U2gp41 CBD-121M785-U1gp41 ID22M785-U223M785-U324M785-U4
31N10-U1 Signal Intensity Values Peptide #Clade AClade BClade CClade DGroup MCRF 1CRF 2335341518958960589785915659086190590705342259159194130651955306774264031213019654161471835128844181350311994341821440230Background subtraction: 5-fold nonspecific peptide binding.Note: “Breakage” of the gp41 ID epitope at peptide #192 and 193 is a knownslide quality issue. The epitope is very likely a continuous one.5 µg/ml, Lot # JPT2049, PMT540; SXS 23-11