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

33 Citations (Scopus)

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

Fingerprint

Immobilized Antibodies
Ethylene Glycol
Polyethylene glycols
Antigens
Glass
Antibodies
Ovalbumin
Bacilli
Bovine Serum Albumin
Immobilization
Heating
Escherichia coli
Bacillus
Enzyme-Linked Immunosorbent Assay
Binding Sites
Molecules

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 journalArticle

@article{48fca5700d31414ba1b6aed5b99a4c2b,
title = "Influence of a poly-ethylene glycol spacer on antigen capture by immobilized antibodies",
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.",
keywords = "Antigen capture, Immobilized antibodies, Spacer",
author = "Weimer, {Bart C} and Walsh, {Marie K.} and Xinwen Wang",
year = "2000",
month = "9",
day = "11",
doi = "10.1016/S0165-022X(00)00114-7",
language = "English (US)",
volume = "45",
pages = "211--219",
journal = "Journal of Proteomics",
issn = "1874-3919",
publisher = "Elsevier",
number = "2",

}

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 -