Heterologous antigen selection of camelid heavy chain single domain antibodies against tetrabromobisphenol A

Jia Wang, Candace R S Bever, Zuzana Majkova, Julie E Dechant, Jun Yang, Shirley J. Gee, Ting Xu, Bruce D. Hammock

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Tetrabromobisphenol A (TBBPA) is a ubiquitous flame retardant. A high-throughput immunoassay would allow for monitoring of human and environmental exposures as a part of risk assessment. Naturally occurring antibodies in camelids that are devoid of light chain, show great promise as an efficient tool in monitoring environmental contaminants, but they have been rarely used for small molecules. An alpaca was immunized with a TBBPA hapten coupled to thyroglobulin and a variable domain of heavy chain antibody (VHH) T3-15 highly selective for TBBPA was isolated from a phage displayed VHH library using heterologous coating antigens. Compared to the VHHs isolated using homologous antigens, VHH T3-15 had about a 10-fold improvement in sensitivity in an immunoassay. This assay, under the optimized conditions of 10% methanol in the assay buffer (pH 7.4), had an IC50 for TBBPA of 0.40 ng mL -1 and negligible cross reactivity (<0.1%) with other tested analogues. After heating the VHH at 90 °C for 90 min about 20% of the affinity for coating antigen T3-BSA remained. The recoveries of TBBPA from spiked soil and fetal bovine serum samples ranged from 90.3% to 110.7% by ELISA and agreed well with a liquid chromatography-tandem mass spectrometry method. We conclude the many advantages of VHH make them attractive for the development of immunoassays to small molecules.

Original languageEnglish (US)
Pages (from-to)8296-8302
Number of pages7
JournalAnalytical Chemistry
Volume86
Issue number16
DOIs
StatePublished - Aug 19 2014

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Single-Domain Antibodies
Heterophile Antigens
Assays
Flame Retardants
CD3 Antigens
Antigens
Coatings
Molecules
Bacteriophages
Antibodies
Haptens
Monitoring
Thyroglobulin
Liquid chromatography
Risk assessment
Mass spectrometry
Methanol
Buffers
Throughput
tetrabromobisphenol A

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Heterologous antigen selection of camelid heavy chain single domain antibodies against tetrabromobisphenol A. / Wang, Jia; Bever, Candace R S; Majkova, Zuzana; Dechant, Julie E; Yang, Jun; Gee, Shirley J.; Xu, Ting; Hammock, Bruce D.

In: Analytical Chemistry, Vol. 86, No. 16, 19.08.2014, p. 8296-8302.

Research output: Contribution to journalArticle

Wang, Jia ; Bever, Candace R S ; Majkova, Zuzana ; Dechant, Julie E ; Yang, Jun ; Gee, Shirley J. ; Xu, Ting ; Hammock, Bruce D. / Heterologous antigen selection of camelid heavy chain single domain antibodies against tetrabromobisphenol A. In: Analytical Chemistry. 2014 ; Vol. 86, No. 16. pp. 8296-8302.
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