Development of dcELISA Method for Rapid Detection of β-conglycinin in Soybean

• Published in: International journal of food engineering Vol. 12; no. 5; pp. 461 - 468
• Main Authors: Zhang, Shiyao, Cao, Kefei, Liu, Dandan, Gaowa, Naren, Bao, Nan, Zhao, Yuan
• Format: Journal Article
• Language: English
• Published: De Gruyter 01.07.2016
• Subjects: allergen; Beta-conglycinin; direct ELISA; Soybean
• ISSN: 2194-5764; 1556-3758
• DOI: 10.1515/ijfe-2015-0359

In this assay, rabbit anti-β-conglycinin polyclonal antibody (Pab) was used as primary antibody and goat anti-rabbit IgG-horseradish peroxidase (HRP) was used as secondary antibody. The optimum incubating time of blocking, HRP and TMB (3, 3’5, 5’-tetramethylbenzidine) detected by the response surface and orthogonal test was 58, 140, and 20 min, respectively. The practical working range for the determination of β-conglycinin was 20–320 ng ml . The recoveries of β-conglycinin in spiked soybean samples were between 95.8 % and 101.2 % with the relative standard deviation less than 6.9 % (intra-assay) and 8.2 % (inter-assay). The current method was used to analyze 80 different soybean obtained from different region of Jilin province and the detected results using the direct ELISA. Compared with the competitive ELISA, The results showed contents had no difference between two methods. The dcELISA assay provides a specific and sensitive method for detecting of soybean β-conglycinin in actual production.