title: A novel interaction mode between acrylamide and its specific antibody.
abstract: Since the discovery of high-level acrylamide (Acr) contamination in food, extensive international studies have focused on its toxicity and detection. By using a novel antigen synthetic strategy, we have successfully obtained a specific antibody towards acrylamide (Acr-Ab). Herein, the Acr-Ab and its interactions with Acr were characterized. Enzyme-linked immunosorbent assay (ELISA) and dynamic light scattering (DLS) investigations revealed that the conformational structure of Acr-Ab was sensitive to buffers. It showed a satisfied immunoreactivity in phosphate buffered saline (PBS), but denatured in water. In natural state, Acr-Ab had a trend of getting aggregation through their complementarity determining regions (CDRs). Adding Acr leaded to their disassembling. While mixed with Acr, Acr-Ab exhibits not only a fast, high-specific, and reversible non covalent binding (by surface plasmon resonance, SPR), but also a covalent alkylation with Acr through cysteine and histidine residues on its surface, as demonstrated by high-performance liquid chromatography (HPLC). Neither of the two reactions involves conformational change in secondary or tertiary structures as shown in circular dichroism spectra (CD). These special properties of Acr-Ab and the entirely new interaction mode with Acr will extend our knowledge of Acr related biosystem and facilitate the development of new detection strategies for Acr.
authors.affiliation: a Beijing National Laboratory for Molecular Sciences (BNLMS) , College of Chemistry and Molecular Engineering, Peking University , Beijing , China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Zhou
authors.forename: Dan
authors.initials: D
authors.lastname: Wang
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.forename: Yunfeng
authors.initials: Y
authors.lastname: Zhao
authors.forename: Meiping
authors.initials: M
authors.lastname: Zhao
authors.forename: Yongning
authors.initials: Y
authors.lastname: Wu
mesh_terms: D020106:Acrylamide / Q000032:analysis / Q000737:chemistry*; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D002942:Circular Dichroism; D004797:Enzyme-Linked Immunosorbent Assay; D005504:Food Analysis / Q000379:methods; D051379:Mice; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D020106:Acrylamide
keywords: acrylamide; alkylation; antibody; immunoaffinity; interaction