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 language | English (US) |
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Pages (from-to) | 211-219 |
Number of pages | 9 |
Journal | Journal of Biochemical and Biophysical Methods |
Volume | 45 |
Issue number | 2 |
DOIs | |
State | Published - Sep 11 2000 |
Externally published | Yes |
Keywords
- Antigen capture
- Immobilized antibodies
- Spacer
ASJC Scopus subject areas
- Biochemistry
- Biophysics