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An indirect competitive enzyme-linked immunosorbent assay for acrylamide detection based on a monoclonal antibody
- Zhu, Yuting, Song, Shanshan, Liu, Liqiang, Kuang, Hua, Xu, Chuanlai
- Food and agricultural immunology 2016 v.27 no.6 pp. 796-805
- acrylamides, antigens, bovine serum albumin, cross reaction, detection limit, enzyme-linked immunosorbent assay, foods, inhibitory concentration 50, monoclonal antibodies, ovalbumin
- Acrylamide (AA) is formed spontaneously in heated foodstuffs and is a focus of concern in many people due to safety. In this study, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on a monoclonal antibody (MAb) to detect a derivative, which was generated from 4-mercaptobenzoic acid (4-MBA). As AA is a very small molecule (71.08 Da) and cannot elicit a homologous monoclonal antibody, we coupled the AA derivative (AA-4-MBA) to a carrier protein such as bovine serum albumin (BSA) and ovalbumin (OVA). The conjugates were used as the immunogen and coating antigen. A rapid and sensitive icELISA against AA-4-MBA was obtained by optimizing the experimental parameters. The MAb which had no specificity for AA or 4-MBA, but had high affinity for AA-4-MBA, had a satisfactory IC ₅₀ of 32 ng/ml and a limit of detection of 8.87 ng/ml. The quantitative working range was 8.87–112.92 ng/ml (IC ₂₀ to IC ₈₀). Cross-reactivity with other analogues was lower than 10%. These results indicated that the developed icELISA was a fast and efficient method for detecting AA in food.