Preparation for aflatoxin B(1)-cationized bovine serum albumin based on Mannich-type reaction.
Aflatoxin B(1) (AFB(1)), which is commonly found in agricultural commodities, is one of the most potent carcinogenic mycotoxins. To ensure food safety, rapid and low-cost immunological methods have been applied to detect AFB(1) worldwide. A key step in these immunological methods is coupling AFB(1) to carrier proteins; AFB(1) is usually deviated to AFB(1)-oxime and coupled to carrier proteins to form the AFB(1)-oxime-protein conjugate. In the current research, AFB(1) was directly coupled with cationized bovine serum albumin (cBSA) using a method based on Mannich-type principles. The coupling effects were investigated with different initial molar ratios of AFB(1) to cBSA. The conjugate molar ratio was 6.4:1 when the initial molar ratio was 40:1. The cationized proteins and their conjugates were identified by UV-Vis and FT-IR spectra, which showed the characteristic bands of the ethylendiamine group and AFB(1), respectively. After BALB/c mice were immunized with AFB(1)-cBSA, a quicker immunological response and a similar sensitivity of antisera against AFB(1) were observed, compared with immunization by AFB(1)-oxime-BSA. This suggests that the Mannich-type reaction might be an alternative method of preparation for AFB(1)-protein.
Zhou
Youxiang
Y
College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China.
Wu
Jiajia
J
Yu
Wei
W
Xu
Ying
Y
Wang
Ping
P
Xie
Bijun
B
Chen
Fusheng
F
D016604:Aflatoxin B1 / Q000737:chemistry* / Q000276:immunology; D000818:Animals; D000918:Antibody Specificity; D002352:Carrier Proteins / Q000737:chemistry* / Q000276:immunology; D002412:Cations; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007158:Immunologic Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D010091:Oximes / Q000737:chemistry* / Q000276:immunology; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000276:immunology
D002352:Carrier Proteins; D002412:Cations; D010091:Oximes; D012710:Serum Albumin, Bovine; D016604:Aflatoxin B1