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Detection of weakly interacting anti‐carbohydrate scFv phages using surface plasmon resonance
Author(s) -
Åström Eva,
Ohlson Sten
Publication year - 2006
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.786
Subject(s) - surface plasmon resonance , phagemid , panning (audio) , phage display , monoclonal antibody , biosensor , conjugate , chemistry , antibody , microbiology and biotechnology , binding selectivity , peptide library , biochemistry , gene , peptide sequence , biology , bacteriophage , peptide , escherichia coli , nanotechnology , materials science , paleontology , mathematical analysis , zoom , mathematics , nanoparticle , immunology , lens (geology)
Abstract Methods to characterise and confirm specificity of scFv displayed on phages are important during panning procedures, especially when selecting for antibody fragments with weak affinities in the millimole to micromole range. In this report the surface plasmon resonance (SPR) biosensor was used to study and verify specificity of phages displaying weak anti‐carbohydrate scFvs. The variable immunoglobulin light (V L ) and heavy (V H ) chain genes of the weak monoclonal antibody 39.5 were amplified and cloned into a phagemid and displayed as a scFv‐pIII fusion protein on filamentous phage. This monoclonal antibody recognises with weak affinity the structural sequence Glc α 1‐4Glc present in a variety of carbohydrate molecules. Injection of the 39.5 phages over a biosensor chip immobilised with a (Glc) 4 ‐BSA conjugate confirmed selective binding of the scFv to its antigen. Inhibition studies verified the specificity. These results clearly show that SPR technology can be used to evaluate in terms of binding and specificity weakly interacting scFv displayed on the phage surface. Copyright © 2006 John Wiley & Sons, Ltd.