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微囊藻毒素-LR完全抗原的设计及制备
引用本文:盛建武,何苗,宋保栋,施汉昌,钱易.微囊藻毒素-LR完全抗原的设计及制备[J].环境科学,2005,26(3):33-37.
作者姓名:盛建武  何苗  宋保栋  施汉昌  钱易
作者单位:清华大学环境科学与工程系,环境模拟与污染控制国家重点联合实验室,北京,100084;清华大学环境科学与工程系,环境模拟与污染控制国家重点联合实验室,北京,100084;清华大学环境科学与工程系,环境模拟与污染控制国家重点联合实验室,北京,100084;清华大学环境科学与工程系,环境模拟与污染控制国家重点联合实验室,北京,100084;清华大学环境科学与工程系,环境模拟与污染控制国家重点联合实验室,北京,100084
基金项目:国家高技术研究发展计划(863)项目(2002AA649160)
摘    要:从偶联位点、偶联剂、载体蛋白和偶联步骤等分析出发,设计了微囊藻毒素-LR (Microcystin-LR ,MC-LR)完全抗原的制备方法.在半抗原分子第7位氨基酸分子上引进1个自由的氨基;再用戊二醛2步法将此中间产物(H2N-etMC-LR)分别与BSA和OVA偶联.中间产物和偶联产物分别经固相萃取和透析纯化后,经SDS凝胶电泳、紫外扫描及生物质谱技术鉴定,结果表明MC-LR与BSA的平均偶联比能达到5以上,满足了进一步免疫的要求.

关 键 词:微囊藻毒素-LR  完全抗原  偶联比
文章编号:0250-3301(2005)03-0033-05
收稿时间:9/6/2004 12:00:00 AM
修稿时间:2004/10/15 0:00:00

Synthesis of Complete Antigen of Microcystin-LR and Its Antibody Production
SHENG Jian-wu,HE Miao,SONG Bao-dong,SHI Han-chang and QIAN Yi.Synthesis of Complete Antigen of Microcystin-LR and Its Antibody Production[J].Chinese Journal of Environmental Science,2005,26(3):33-37.
Authors:SHENG Jian-wu  HE Miao  SONG Bao-dong  SHI Han-chang and QIAN Yi
Institution:ESPC State Key Joint Laboratory, Department of Environment Science and Engineering, Tsinghua University, Beijing 100084, China.
Abstract:Based on the analysis of coupling location, coupling regent, carrying protein and coupling process, completed antigen of Microcystin-LR (MC-LR) was synthesized. A free amidogen was introduced to microcystin-LR (MC-LR) using chemical modification (aminoethylation) of its 7th core amino acids, N-methyl-dehydroalanine. The intermediate product Aminoethyl-MC-LR (H2N-etMC-LR) was purified by Solid Phase Elute (SPE) and identified by Mass Spectrometry,then coupled with BSA through glutaraldehyde to be complete antigen. The completed antigen was dialyzed and identified by SDS-PAGE,UV-scanning and bio-mass spectrometry. The result show that the coupling ratio between MC-LR and BSA of complete antigen in average was over 5, and it is sufficient to immunity.
Keywords:Microcystin-LR  complete antigen  coupling ratio
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