Magnetic bead enzyme-linked immunosorbent assay (ELISA) detects antigen-specific binding by phage-displayed scFv antibodies that are not detected with conventional ELISA

Analytical Biochemistry
M KalaS Sinha

Abstract

An efficient means for the detection of antigen-specific binding by phage-displayed antibodies would facilitate the selection of such phage, especially from libraries with large repertoires of V-genes. We report the development and characterization of a magnetic bead phage ELISA which detects antigen binding phage which could not be detected by conventional ELISA. We were attempting to select phage binding to the oncodevelopmental antigen, heat-stable alkaline phosphatase (HSAP). Although there was an obvious enrichment in the phage titers after successive rounds of selection, we were unable to detect antigen-binding phage by ELISA on a plastic surface. However, ELISA with a suspension of superparamagnetic particles covalently conjugated to HSAP effectively identified antigen-binding phage after the fourth round of selection. This method could also detect antigen-specific binding of individual phage clones. Some of the phage clones bound to either amino- or carboxy-terminal-conjugated HSAP, perhaps reflecting the differences in the exposed epitopes. It is suggested that a sensitive method such as magnetic bead phage ELISA be tried before declaring a phage selection as unsuccessful or concluding that a phage clone does not bind ...Continue Reading

References

Aug 5, 1992·Journal of Molecular Biology·R E HawkinsG Winter
Sep 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·C F BarbasS J Benkovic
Jan 1, 1990·Urological Research·K Koshida, B Wahren
Apr 1, 1990·Clinical Biochemistry·W H Fishman
May 1, 1990·Cell and Tissue Research·E J NouwenM E De Broe
Jul 11, 1995·Nucleic Acids Research·G A Kingsbury, R P Junghans

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Citations

Oct 28, 2009·Hybridoma·Pooja GoswamiSubrata Sinha
Apr 13, 2010·Journal of Immunological Methods·Francisco Bruno Moreira Brás GomesJoão Paulo Medeiros Ferreira
Aug 15, 2006·Journal of Immunological Methods·Brigitte SimonsMary Alice Hefford
Oct 17, 2014·Journal of Immunoassay & Immunochemistry·Mukesh KumarSubrata Sinha
Sep 1, 2016·Scientific Reports·Yi-Hsing HsiaoChihchen Chen
Aug 10, 2018·Current Protocols in Protein Science·Heungnam Kim, Mitchell Ho
Jul 7, 2017·Cold Spring Harbor Protocols·Thomas O Kohl, Carl A Ascoli

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