Phage display selection of EGFR-specific antibodies by capture-sandwich panning

Min Kyung Ki, Kyung Jae Kang, Hyunbo Shim

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Sandwich ELISA experiment requires two mutually compatible affinity reagents, typically antibodies, that are highly sensitive and specific to the target analyte in its native conformation, and whose epitopes do not overlap each other's. Finding or developing a pair of antibodies that meet these requirements can be a challenge and many otherwise useful antibodies fail in sandwich ELISA-format analysis. In order to discover sandwich immunoassay-compatible antibodies for epidermal growth factor receptor (EGFR), we first immobilized cetuximab, a high-affinity monoclonal antibody against EGFR, on a plastic surface and affinity-captured EGFR in A431 cell lysate. Panning of a phage antibody library on the surface-captured antigen yielded a couple of antibody clones that can be paired with cetuximab and also with each other in the sandwich-format immunoassay. This provides a potentially useful strategy for the development of sandwich ELISA reagents or antibodies against antigens that are hard to purify.

Original languageEnglish
Pages (from-to)152-156
Number of pages5
JournalBiotechnology and Bioprocess Engineering
Volume15
Issue number1
DOIs
StatePublished - Feb 2010

Keywords

  • Antibody
  • Capture-sandwich panning
  • Epidermal growth factor receptor (EGFR)
  • Phage display
  • Sandich ELISA

Fingerprint

Dive into the research topics of 'Phage display selection of EGFR-specific antibodies by capture-sandwich panning'. Together they form a unique fingerprint.

Cite this