A novel interaction mode between acrylamide and its specific antibody.

A novel interaction mode between acrylamide and its specific antibody. J Immunoassay Immunochem. 2015;36(3):295-311 Authors: Zhou S, Wang D, Zhang C, Zhao Y, Zhao M, Wu Y 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 in...
Source: Journal of Immunoassay and Immunochemistry - Category: Biochemistry Tags: J Immunoassay Immunochem Source Type: research