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

Mrinalini Kala, Kiran Bajaj, Subrata Sinha

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

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 antigen.

Original languageEnglish (US)
Pages (from-to)263-266
Number of pages4
JournalAnalytical Biochemistry
Volume254
Issue number2
DOIs
StatePublished - Dec 15 1997
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Magnetic bead enzyme-linked immunosorbent assay (ELISA) detects antigen- specific binding by phage-displayed scFv antibodies that are not detected with conventional ELISA'. Together they form a unique fingerprint.

Cite this