ITEM-THREE analysis of a monoclonal anti-malaria antibody reveals its assembled epitope on the pfMSP119 antigen

Kwabena F.M. Opuni, Cornelia Koy, Manuela Russ, Maren Reepmeyer, Bright D. Danquah, Moritz Weresow, Astrid Alef, Peter Lorenz, Hans Juergen Thiesen, Michael O. Glocker

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

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 antibody's epitope on the respective antigen. We have cloned a malarial antigen-containing fusion protein, MBP-pfMSP119, in Escherichia coli, which then was structurally and functionally characterized before and after high pressure-assisted enzymatic digestion. We then used our previously developed method, intact transition epitope mapping-targeted high-energy rupture of extracted epitopes (ITEM-THREE), to map the area on the MBPpfMSP119 antigen surface that is recognized by the antipfMSP119 antibody G17.12. We identified three epitope-carrying peptides, 386GRNISQHQCVKKQCPQNSGCFRHLDE411386GRNISQHQCVKKQCPQNSGCFRHLDEREE414, and 415CKC-LLNYKQE424, from the GluC-derived peptide mixture. These peptides belong to an assembled (conformational) epitope on the MBP-pfMSP119 antigen whose identification was substantiated by positive and negative control experiments. In conclusion, our data help to establish a workflow to obtain high-quality control data for diagnostic assays, including the use of ITEM-THREE as a powerful analytical tool. Data are available via ProteomeXchange: PXD019717.

Original languageEnglish
Pages (from-to)14987-14997
Number of pages11
JournalJournal of Biological Chemistry
Volume295
Issue number44
DOIs
Publication statusPublished - 30 Oct 2020

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