Epitope mapping of an anti-plasmodium pfMSP119 antibody by Intact Transition Epitope Mapping - Targeted High-Energy Rupture of Extracted Epitopes

ABSTRACT: 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.

INSTRUMENT(S): Bruker Daltonics flex series, Synapt MS

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Michael Kreutzer  

LAB HEAD: Prof. Dr. Michael O. Glocker

PROVIDER: PXD019717 | Pride | 2020-08-24

REPOSITORIES: Pride