Generation of a novel affibody molecule targeting Chlamydia trachomatis MOMP

Chlamydia trachomatis (C. trachomatis) is the leading cause of preventable blindness worldwide and the most prevalent cause of bacterial sexually transmitted diseases. At present, there is no available vaccine, and recurrences after antibiotics treatment are substantial problems. Major outer membrane protein (MOMP) accounts for 60% of the outer mass of C. trachomatis, functioning as trimeric porin, and it is highly antigenic. Therefore, MOMP is the most promising candidate for vaccine developing and target therapy of Chlamydia. Affibody, a new class of affinity ligands derived from the Z-domain in the binding region of Staphylococcus aureus protein A, has been the focus of researchers as a viable alternative to antibodies. In this study, the MOMP-targeted affibody molecule (Z_MOMP:461) was screened by phage-displayed peptide library. Further, the affinity and specificity were characterized by surface plasmon resonance (SPR) and Western blot. Immunofluorescence assay (IFA) indicated that the MOMP-binding affibody could recognize native MOMP in HeLa229 cells infected C. trachomatis. Immunoprecipitation assay confirmed further that Z_MOMP:461 molecule specifically recognizes the epitope on relaxed trimer MOMP. Our findings provide strong evidence that affibody molecule (Z_MOMP:461) serves as substitute for MOMP antibody for biological applications and has a great potential for delivering drugs for target therapy. KEY POINTS : • We screened a novel affibody molecule Z_MOMP:461 targeting Chlamydia trachomatis MOMP. • Z_MOMP:461 recognizes the recombinant and native MOMP with high affinity and specificity. • Z_MOMP:461 could be internalized into live target cells.

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