Development of enantioselective polyclonal antibodies to detect styrene oxide protein adducts

Shuijie Shen, Fan Zhang, Su Zeng, Ye Tian, Xiaojuan Chai, Shirley Gee, Bruce D. Hammock, Jiang Zheng

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Styrene has been reported to be pneumotoxic and hepatotoxic in humans and animals. Styrene oxide, a major reactive metabolite of styrene, has been found to form covalent binding with proteins, such as albumin and hemoglobin. Styrene oxide has two optical isomers and it was reported that the (R)-enantiomer was more toxic than the (S)-enantiomer. The purpose of this study was to develop polyclonal antibodies that can stereoselectively recognize proteins modified by styrene oxide enantiomers at cysteine residues. Immunogens were prepared by alkylation of thiolated keyhole limpet hemocyanin (KLH) with styrene oxide enantiomers. Polyclonal antibodies were raised by immunization of rabbits with the chiral immunogens. Titration tests showed all six rabbits generated high titers of antisera that recognize (R)- or (S)- coating antigens accordingly. No cross-reaction was observed toward the carrier protein (BSA). All three rabbits immunized with (R)-immunogen produced antibodies that show enantioselectivity to the corresponding antigen, while only one among the three rabbits immunized with (S)-immunogen generated antibodies with enantioselectivity of the recognition. The enantioselectivity was also observed in competitive ELISA and immunoblot analysis. Additionally, competitive ELISA tests showed that the immunorecognition required the hydroxyl group of the haptens. Immunoblot analysis demonstrated that the immunorecognition depended on the amount of protein adducts blotted and hapten loading in protein adducts. In summary, we successfully developed polyclonal antibodies to stereoselectively detect protein adducts modified by styrene oxide enantiomers.

Original languageEnglish (US)
Pages (from-to)2668-2677
Number of pages10
JournalAnalytical Chemistry
Volume81
Issue number7
DOIs
StatePublished - Apr 1 2009

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styrene oxide
Enantiomers
Enantioselectivity
Antibodies
Styrene
Haptens
Proteins
Immunization
Antigens
Poisons
Alkylation
Metabolites
Titration
Isomers
Hydroxyl Radical
Cysteine
Immune Sera
Albumins
Carrier Proteins
Hemoglobins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Shen, S., Zhang, F., Zeng, S., Tian, Y., Chai, X., Gee, S., ... Zheng, J. (2009). Development of enantioselective polyclonal antibodies to detect styrene oxide protein adducts. Analytical Chemistry, 81(7), 2668-2677. https://doi.org/10.1021/ac8023262

Development of enantioselective polyclonal antibodies to detect styrene oxide protein adducts. / Shen, Shuijie; Zhang, Fan; Zeng, Su; Tian, Ye; Chai, Xiaojuan; Gee, Shirley; Hammock, Bruce D.; Zheng, Jiang.

In: Analytical Chemistry, Vol. 81, No. 7, 01.04.2009, p. 2668-2677.

Research output: Contribution to journalArticle

Shen, S, Zhang, F, Zeng, S, Tian, Y, Chai, X, Gee, S, Hammock, BD & Zheng, J 2009, 'Development of enantioselective polyclonal antibodies to detect styrene oxide protein adducts', Analytical Chemistry, vol. 81, no. 7, pp. 2668-2677. https://doi.org/10.1021/ac8023262
Shen, Shuijie ; Zhang, Fan ; Zeng, Su ; Tian, Ye ; Chai, Xiaojuan ; Gee, Shirley ; Hammock, Bruce D. ; Zheng, Jiang. / Development of enantioselective polyclonal antibodies to detect styrene oxide protein adducts. In: Analytical Chemistry. 2009 ; Vol. 81, No. 7. pp. 2668-2677.
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