Project description:Epitope mapping of 4 monoclonal antibodies directed against Neisserial Heparin Binding Antigen by peptide microarray
| PRJNA386498 | ENA
Project description:Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen using next generation sequencing of antigen-specific libraries
Project description:The aim of the study was to determine the epitope targeted by 31E10/E7 mouse monoclonal antibody (mAb) and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. mAb 31E10/E7 was diluted at 1:200 and incubated on a custom PepStar Peptide Microarray platform printed with 560 different peptides in three independent experiments.
Project description:The aim of the study was to determine the epitope targeted by four different HumAbs and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. the HumAbs were diluted at 1:60 and incubated on a custom PepStar Peptide Microarray platform printed with 561 different peptides.
Project description:Rapid diagnostic tests are first line assays for diagnosing infectious diseases, such as malaria. To minimize false positive and false negative test results in population screening assays, high quality reagents and well characterized antigens and antibodies are needed. An important property of antigen - antibody binding is recognition specificity which best can be estimated by mapping an antibodys epitope. The MBP-pfMSP119 antigen was chosen as mutual target for population screening. Also, since an anti-pfMSP119 antibody is planned to function as positive control in screening assays, its binding characteristics to the antigen was investigated. Intact Transition Epitope Mapping - Targeted High-Energy Rupture of Extracted Epitopes (ITEM-THREE) was carried out to map the epitope of a monoclonal anti-pfMSP119 antibody, i.e. the recognized area on the MBP-pfMSP119 antigen surface. The MBP-pfMSP119 fusion protein was cloned and expressed in E.coli which then was enriched by affinity purification on amylose resin. The enriched and purified MBP-pfMSP119 fusion protein was structurally and functionally characterized before and after high pressure-assisted tryptic digestion or after GluC digestion of the reduced and alkylated fusion protein.
