[Conjugation and immunogenic evaluation of complete immunogen for the small molecular environmental pollutants 2,4-D].
Conjugation of complete immunogen for 2,4-dichlorophenoxyacetic acid (2,4-D) was studied. 2,4-D was cross-linked to bovine serum albumin (BSA, carrier) by 1-ethyl-3-(3-dimethyl- aminopropyl) carbodiimide hydrochloride (EDC). The conjugation reaction was found to be more effective at 4 degrees C and incubated for 18 hour. 2,4-D was dissolved in 0.05 mol/L phosphate buffer between 10.0 - 12.0 mg/mL concentration, pH was adjusted to be 5.4 - 6.1. When the weight of added EDC was below 12 mg, the more EDC was added the higher substitution degree complete immunogen was synthesized. Complete immunogen of various substitution degree (2,4-D: protein) were applied to immunize balb/c mice. The conjugates of 2,4-D and poly-L-lysine was applied as coating antigen. It was experimentally found that complete immunogen of substitution degree 12 and 18 are more immunogenic than that of substitution degree 6 and 25. None-specific adsorption between antiserum that was produced by complete immunogen of substitution degree 18 and coating antigen was very weak, and the antiserum contained more 2,4-D specific antibody. It could be used as the immunogen for the preparation monoclonal antibody.
Yu
Ruo-Zhen
RZ
Environmental Simulation and Pollutants Control State Key Joint Laboratory, Department of Environmental Science and Engineering, Tsinghua University, Beijing 100084, China. yuruozheng00@mails.tsinghua.edu.cn
He
Miao
M
Shi
Han-Chang
HC
Liu
Ju
J
Guo
Jie-Han
JH
Wang
Na
N
Qian
Yi
Y
D015084:2,4-Dichlorophenoxyacetic Acid / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000941:Antigens / Q000096:biosynthesis / Q000276:immunology; D003432:Cross-Linking Reagents / Q000737:chemistry; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives* / Q000737:chemistry; D006540:Herbicides / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine
D000911:Antibodies, Monoclonal; D000941:Antigens; D003432:Cross-Linking Reagents; D006540:Herbicides; D015084:2,4-Dichlorophenoxyacetic Acid; D012710:Serum Albumin, Bovine; C041829:1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide methiodide; D005022:Ethyldimethylaminopropyl Carbodiimide