Development and utilization of camelid VHH antibodies from alpaca for 2,2′,4,4′-tetrabrominated diphenyl ether detection

Candace R S Bever, Zuzana Majkova, Rajeswaran Radhakrishnan, Ian Suni, Mark McCoy, Yanru Wang, Julie E Dechant, Shirley Gee, Bruce D. Hammock

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Abstract

An antibody-based analytical method for the detection of a chemical flame retardant using antibody fragments isolated from an alpaca has been developed. One specific chemical flame retardant congener, 2,2′,4,4′- tetrabrominated diphenyl ether (BDE-47), is often the major poly-BDE (PBDE) congener present in human and environmental samples and that which is the most frequently detected. An alpaca was immunized with a surrogate of BDE-47 covalently attached to a carrier protein. The resulting mRNA coding for the variable domain of heavy-chain antibodies (VHH) were isolated, transcribed to cDNA, and cloned into a phagemid vector for phage display library construction. Selection of VHHs recognizing BDE-47 was achieved by panning under carefully modified conditions. The assay sensitivity for detecting BDE-47 was down to the part-per-billion (microgram per liter) level. Cross-reactivity analyses confirmed that this method was highly selective for BDE-47 and selected hydroxylated metabolites. When exposed to elevated temperatures, the camelid VHH antibodies retained more reactivity than a polyclonal antibody developed to the same target analyte. The use of this VHH antibody reagent immobilized onto a Au electrode for impedance biosensing demonstrates the increased versatility of VHH antibodies.

Original languageEnglish (US)
Pages (from-to)7875-7882
Number of pages8
JournalAnalytical Chemistry
Volume86
Issue number15
DOIs
StatePublished - Aug 5 2014

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ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Bever, C. R. S., Majkova, Z., Radhakrishnan, R., Suni, I., McCoy, M., Wang, Y., Dechant, J. E., Gee, S., & Hammock, B. D. (2014). Development and utilization of camelid VHH antibodies from alpaca for 2,2′,4,4′-tetrabrominated diphenyl ether detection. Analytical Chemistry, 86(15), 7875-7882. https://doi.org/10.1021/ac501807j