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) |
|---|---|
| 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 |
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Keywords
- Antigen capture
- Immobilized antibodies
- Spacer
ASJC Scopus subject areas
- Biochemistry
- Biophysics
Cite this
Influence of a poly-ethylene glycol spacer on antigen capture by immobilized antibodies. / Weimer, Bart C; Walsh, Marie K.; Wang, Xinwen.
In: Journal of Biochemical and Biophysical Methods, Vol. 45, No. 2, 11.09.2000, p. 211-219.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Influence of a poly-ethylene glycol spacer on antigen capture by immobilized antibodies
AU - Weimer, Bart C
AU - Walsh, Marie K.
AU - Wang, Xinwen
PY - 2000/9/11
Y1 - 2000/9/11
N2 - 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.
AB - 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.
KW - Antigen capture
KW - Immobilized antibodies
KW - Spacer
UR - http://www.scopus.com/inward/record.url?scp=0034638028&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034638028&partnerID=8YFLogxK
U2 - 10.1016/S0165-022X(00)00114-7
DO - 10.1016/S0165-022X(00)00114-7
M3 - Article
C2 - 10989137
AN - SCOPUS:0034638028
VL - 45
SP - 211
EP - 219
JO - Journal of Proteomics
JF - Journal of Proteomics
SN - 1874-3919
IS - 2
ER -