Comparison of antibody functionality using different immobilization methods

R. Danczyk, B. Krieder, A. North, T. Webster, H. HogenEsch, A. Rundell

Research output: Contribution to journalArticle

103 Scopus citations

Abstract

This study investigates the influence of antibody immobilization methods on antigen capture. Adsorption and two surface chemistries, an aminosilane chemistry and a common heterobifunctional crosslinker (N-gamma-maleimidobutyryloxy-succinimide ester, GMBS), were compared and evaluated for their ability to immobilize antibodies and capture antigen. The role of protein A as an orienting protein scaffold component in each of these techniques was also evaluated. Through experimentation it was determined that the GMBS technique immobilized the highest amount of antibody and minimized nonspecific binding. For all techniques, the most functional antibodies were found to be those immobilized with protein A. Interestingly, the aminosilane technique demonstrated the highest antigen capture with antibody alone but also exhibited the highest level of nonspecific binding.

Original languageEnglish (US)
Pages (from-to)215-223
Number of pages9
JournalBiotechnology and Bioengineering
Volume84
Issue number2
DOIs
StatePublished - Oct 20 2003
Externally publishedYes

Keywords

  • Antibody
  • Antigen capture
  • Biosensor
  • Protein A
  • Protein immobilization

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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