Influence of a poly-ethylene glycol spacer on antigen capture by immobilized antibodies

Bart C Weimer, Marie K. Walsh, Xinwen Wang

Research output: Contribution to journalArticle

35 Scopus citations

Abstract

The use of spacers to distance an immobilized antibody from the surface of a support matrix introduces flexibility, which can reduce steric interferences between antibodies leading to a higher antigen capture efficiency. In this paper we investigated the use of a spacer molecule, poly- ethylene glycol (PEG), between the matrix surface and antibodies for the capture of Bacillus globigii, E. coli 0157:H7, and ovalbumin. The antigen capture efficiency was determined using a surface ELISA method. Antibodies against the antigens were covalently immobilized either directly or via PEG to glass surfaces using a one-step EDC reaction. The amount of antibody immobilized was determined before blocking the nonspecific binding sites with bovine serum albumin. Antibodies immobilized via a PEG spacer showed a higher capture efficiency compared to direct immobilization, which was more pronounced with large antigens. Antibodies immobilized on glass supports were stable at 65°C for at least 80 min, and the capture efficiency increased with heating at 65°C for 20 min. (C) 2000 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)211-219
Number of pages9
JournalJournal of Biochemical and Biophysical Methods
Volume45
Issue number2
DOIs
StatePublished - Sep 11 2000
Externally publishedYes

Keywords

  • Antigen capture
  • Immobilized antibodies
  • Spacer

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

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