Exploring epitopes of antibodies toward the human tryptophanyl-tRNA synthetase

Hjelm, Barbara (author): KTH,Albanova VinnExcellence Center for Protein Technology, ProNova

Fernandez, Carmen Diez (author): KTH,Albanova VinnExcellence Center for Protein Technology, ProNova

Ståhl, Stefan (author): KTH,Albanova VinnExcellence Center for Protein Technology, ProNova

Johannesson, Henrik (author): KTH,Albanova VinnExcellence Center for Protein Technology, ProNova

Rockberg, Johan (author): KTH,Albanova VinnExcellence Center for Protein Technology, ProNova

(creator_code:org_t)

Related links:: https://urn.kb.se/re...; show more...; https://doi.org/10.1...; show less...

Abstract Subject headings

There is a need to characterize the epitopes of affinity reagents to develop high quality affinity reagents for research, diagnostics and therapy. Here, we describe the analysis of epitopes of antibodies generated toward human tryptophanyl-tRNA synthetase (WARS) using both combinatorial bacterial display and suspension bead array. The bacterial display revealed that the polyclonal antibody binds to three separate epitopes and peptide scanning using 15-mers revealed binding to a 13 amino acid consensus sequence (ELINRIERATGQR). A mouse monoclonal antibody was generated and the mapping approach revealed binding toward a slightly shifted position of the same epitope. Structural analysis showed that the antibodies bind to a-helical regions on the surface of the target protein. An alanine-scanning experiment showed binding to four specific residues. The implications for the systematic analysis of antibody epitopes on the basis of these results are discussed.

By the author/editor: Hjelm, Barbara; Fernandez, Carme ...; Löfblom, John; Ståhl, Stefan; Johannesson, Hen ...; Rockberg, Johan; show more...; Uhlen, Mathias; show less...

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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