Novel artificial antigen synthesis for antibody production and development of an indirect competitive ELISA of cyanocobalamin
Novel artificial antigen synthesis for antibody production and development of an indirect competitive ELISA of cyanocobalamin
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用于抗体生产的新型人工抗原合成以及氰钴胺素间接竞争 ELISA 的开发
DOI：
10.1039/c5ay00995b
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发表时间：
2015-06
影响因子：
3.1
通讯作者：
Liu Renrong
中科院分区：
文献类型：
--
作者：
Yang Fanfan;Zhu Lixin;Meng Wei;Liu Renrong
在当前的工作中，氰钴胺通过碳二亚胺（CDI）法与牛血清白蛋白（BSA）和卵清蛋白（OVA）偶联以制备人工抗原。然后获得了一种针对氰钴胺的单克隆抗体（MAB）。与其他已报道的抗体相比……
In the current work, cyanocobalamin is coupled to Bovine Serum Albumin (BSA) and ovalbumin (OVA) by a CDI method to produce artificial antigens. Then a monoclonal antibody (MAB) against cyanocobalamin was acquired. In comparison with other reported antibodies, the MAB showed good specificity toward cyanocobalamin. Cross reactivity was found to be less than 0.01% with other compounds of vitamin B, except for three analogs of cobalamins which showed a cross reactivity from 0.27% to 2.31%. An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for the detection of cyanocobalamin was developed based on the MAB. Under the optimum conditions, the limit of detection was 0.2 ng mL−1 with a linear working range of 2–100 ng mL−1 (R2 = 0.993). The spiked samples were detected with recovery ranging from 86.02–110.54%, and the coefficients of variation were from 2.62% to 13.7%. Six commercially available vitamin supplements were tested by the ELISA method, after purification with an immunoaffinity column, the samples were validated by HPLC with UV detection at 361 nm. The results obtained also showed good correlation between ELISA and HPLC (R2 = 0.998). Thus, the method proposed had been proved to be a trustworthy tool to quantify cyanocobalamin in vB12 tablet supplements.
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影响因子：
1.7
作者：
Ni Zhang;W. Wang;Kuigang Wang;Zhiying Ding;Zeng Wei
通讯作者：
Ni Zhang;W. Wang;Kuigang Wang;Zhiying Ding;Zeng Wei
影响因子：
--
作者：
A. Molloy;J. Scott
通讯作者：
A. Molloy;J. Scott
影响因子：
--
作者：
C. Ju;Youhua Xiong;A. Gao;Tang-bin Yang;Lei Wang
通讯作者：
C. Ju;Youhua Xiong;A. Gao;Tang-bin Yang;Lei Wang
影响因子：
5.9
作者：
Watanabe F;Yabuta Y;Bito T;Teng F
通讯作者：
Teng F
影响因子：
1.1
作者：
N. Akbay;E. Gök
通讯作者：
N. Akbay;E. Gök