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Citation: ZHANG Ming-Cui, ZHUANG Hui-Sheng, LANG Qing. Immunological Assaying of Environmental Hormone Dibutyl Phthalate Ⅱ——Preparation of Antibody[J]. Chinese Journal of Applied Chemistry, ;2007, 24(5): 530-533. shu

Immunological Assaying of Environmental Hormone Dibutyl Phthalate Ⅱ——Preparation of Antibody

1.
a College of Environmental Science and Engineering, Donghua University, Shanghai 200051;
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b College of Chemistry and Materials Science, Anhui Normal University, Wuhu

Corresponding author: ZHUANG Hui-Sheng,
Received Date: 23 August 2006
Available Online: 23 November 2006
Fund Project: 国家自然科学基金资助项目(20277005) (20277005)

The haptene of dibutyl phthalate (DBP) was conjugated to bovine serum by azo coupling.The conjugate was used as an immunogen.High titer polyclonal antibodies against dibutyl phthalate were produced from New Zealand white rabbits immunized with the immunogen.The final sera had titers against DBP with dilutions of 1:32~1:64 with double immunodiffusion,and 1:6 400~1:12 800 with indirect competitive fluorescence immunoassay.After an acceptable antibody titer was observed,the animals were exsanguinated and antiserum was purified.The purified IgG fractions were lyophilized aliquoted into vials,and stored at -20℃.An indirect fluorescence immunoassay to detect antibody was developed.The analysis experiment of anti-DBP by indirect competitive fluorescence immunoassay showed that anti-DBP antibody was crossreacted with other similar phthalate compounds,such as dimethyl phthalate (DMP),diethyl phthalate (DEP),dipropyl phthalate (DPrP),dicyclohexyl phthalate (DCHP),di-n-pentyl phthalate (DPP) and diethylhexyl phthalate (DEHP),and the cross reactivities were less than 10%.Therefore,this anti-DBP antibody could be interesting for developing the fluorescence immunoassay of DBP.
- dibutyl phthalate,
- antibody,
- preparation,
- fluorescence immunoassay
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