Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis

Hassan K M Bekheit, Anne D. Lucas, Shirley J. Gee, Robert O. Harrison, Bruce D. Hammock

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of β-exotoxin from Bacillus thuringiensis in solution and to evaluate the ability of the antibodies to distinguish among various natural and synthetic compounds related to the β-exotoxin. The β-exotoxin was coupled to several proteins via glutaraldehyde, diazotization, and periodate procedures. The antibodies used in the assay were obtained from antisera raised against β-exotoxin linked to keyhole limpet hemocyanin, and the ELISA coating antigens consisted of β-exotoxin-bovine serum albumin conjugates. Both homologous and heterologous ELISA systems were examined. The homologous systems were not useful because the free β-exotoxin did not inhibit antibody binding to the solid phase. The heterologous systems yielded the most sensitive assays, but antisera obtained from all of the immunogens were used successfully in developing ELISAs for β-exotoxin. With these sensitive ELISAs, β-exotoxin was detected in samples of commercial formulations at levels as low as 0.1 ng/mL. A good correlation was observed when culture media was fortified with β-exotoxin and analyzed in a blind fashion with both HPLC and ELISA. These data suggest that the ELISA could be a valuable tool for detecting and quantifying β-exotoxin.

Original languageEnglish (US)
Pages (from-to)1530-1536
Number of pages7
JournalJournal of Agricultural and Food Chemistry®
Volume41
Issue number9
StatePublished - 1993

Fingerprint

exotoxins
Exotoxins
Immunosorbents
Bacillus thuringiensis
Bacilli
Assays
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
Enzymes
antibodies
antiserum
Antibodies
Immune Sera
antigens
glutaraldehyde
Glutaral
assays
bovine serum albumin
Bovine Serum Albumin
coatings

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)

Cite this

Bekheit, H. K. M., Lucas, A. D., Gee, S. J., Harrison, R. O., & Hammock, B. D. (1993). Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis. Journal of Agricultural and Food Chemistry®, 41(9), 1530-1536.

Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis. / Bekheit, Hassan K M; Lucas, Anne D.; Gee, Shirley J.; Harrison, Robert O.; Hammock, Bruce D.

In: Journal of Agricultural and Food Chemistry®, Vol. 41, No. 9, 1993, p. 1530-1536.

Research output: Contribution to journalArticle

Bekheit, HKM, Lucas, AD, Gee, SJ, Harrison, RO & Hammock, BD 1993, 'Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis', Journal of Agricultural and Food Chemistry®, vol. 41, no. 9, pp. 1530-1536.
Bekheit, Hassan K M ; Lucas, Anne D. ; Gee, Shirley J. ; Harrison, Robert O. ; Hammock, Bruce D. / Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis. In: Journal of Agricultural and Food Chemistry®. 1993 ; Vol. 41, No. 9. pp. 1530-1536.
@article{e744507bc7c947ddbc4479310b2b9758,
title = "Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis",
abstract = "A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of β-exotoxin from Bacillus thuringiensis in solution and to evaluate the ability of the antibodies to distinguish among various natural and synthetic compounds related to the β-exotoxin. The β-exotoxin was coupled to several proteins via glutaraldehyde, diazotization, and periodate procedures. The antibodies used in the assay were obtained from antisera raised against β-exotoxin linked to keyhole limpet hemocyanin, and the ELISA coating antigens consisted of β-exotoxin-bovine serum albumin conjugates. Both homologous and heterologous ELISA systems were examined. The homologous systems were not useful because the free β-exotoxin did not inhibit antibody binding to the solid phase. The heterologous systems yielded the most sensitive assays, but antisera obtained from all of the immunogens were used successfully in developing ELISAs for β-exotoxin. With these sensitive ELISAs, β-exotoxin was detected in samples of commercial formulations at levels as low as 0.1 ng/mL. A good correlation was observed when culture media was fortified with β-exotoxin and analyzed in a blind fashion with both HPLC and ELISA. These data suggest that the ELISA could be a valuable tool for detecting and quantifying β-exotoxin.",
author = "Bekheit, {Hassan K M} and Lucas, {Anne D.} and Gee, {Shirley J.} and Harrison, {Robert O.} and Hammock, {Bruce D.}",
year = "1993",
language = "English (US)",
volume = "41",
pages = "1530--1536",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society",
number = "9",

}

TY - JOUR

T1 - Development of an enzyme-linked immunosorbent assay for the β-exotoxin of bacillus thuringiensis

AU - Bekheit, Hassan K M

AU - Lucas, Anne D.

AU - Gee, Shirley J.

AU - Harrison, Robert O.

AU - Hammock, Bruce D.

PY - 1993

Y1 - 1993

N2 - A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of β-exotoxin from Bacillus thuringiensis in solution and to evaluate the ability of the antibodies to distinguish among various natural and synthetic compounds related to the β-exotoxin. The β-exotoxin was coupled to several proteins via glutaraldehyde, diazotization, and periodate procedures. The antibodies used in the assay were obtained from antisera raised against β-exotoxin linked to keyhole limpet hemocyanin, and the ELISA coating antigens consisted of β-exotoxin-bovine serum albumin conjugates. Both homologous and heterologous ELISA systems were examined. The homologous systems were not useful because the free β-exotoxin did not inhibit antibody binding to the solid phase. The heterologous systems yielded the most sensitive assays, but antisera obtained from all of the immunogens were used successfully in developing ELISAs for β-exotoxin. With these sensitive ELISAs, β-exotoxin was detected in samples of commercial formulations at levels as low as 0.1 ng/mL. A good correlation was observed when culture media was fortified with β-exotoxin and analyzed in a blind fashion with both HPLC and ELISA. These data suggest that the ELISA could be a valuable tool for detecting and quantifying β-exotoxin.

AB - A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify the amount of β-exotoxin from Bacillus thuringiensis in solution and to evaluate the ability of the antibodies to distinguish among various natural and synthetic compounds related to the β-exotoxin. The β-exotoxin was coupled to several proteins via glutaraldehyde, diazotization, and periodate procedures. The antibodies used in the assay were obtained from antisera raised against β-exotoxin linked to keyhole limpet hemocyanin, and the ELISA coating antigens consisted of β-exotoxin-bovine serum albumin conjugates. Both homologous and heterologous ELISA systems were examined. The homologous systems were not useful because the free β-exotoxin did not inhibit antibody binding to the solid phase. The heterologous systems yielded the most sensitive assays, but antisera obtained from all of the immunogens were used successfully in developing ELISAs for β-exotoxin. With these sensitive ELISAs, β-exotoxin was detected in samples of commercial formulations at levels as low as 0.1 ng/mL. A good correlation was observed when culture media was fortified with β-exotoxin and analyzed in a blind fashion with both HPLC and ELISA. These data suggest that the ELISA could be a valuable tool for detecting and quantifying β-exotoxin.

UR - http://www.scopus.com/inward/record.url?scp=0000662325&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0000662325&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:0000662325

VL - 41

SP - 1530

EP - 1536

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 9

ER -