新型抗苯并[a]芘人源单链抗独特型抗体
New human single chain anti-idiotypic antibody against benzo[a]pyrene.
作者信息
Studennikov Artem E, Ustinov Valentin A, Morozova Vera V, Tikunova Nina V, Glushkov Andrey N
机构信息
Federal State Scientific Institute, Federal Research Centre for Coal and Coal Chemistry, Siberian Branch of the Russian Academy of Sciences, Institute of Human Ecology, Kemerovo, Russia.
Federal State Scientific Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
出版信息
Cent Eur J Immunol. 2017;42(2):123-130. doi: 10.5114/ceji.2017.69353. Epub 2017 Aug 8.
The nal¨ve library from the lymphocytes of healthy humans was screened by murine single-stranded idiotypic antibodies against benzo[a]pyrene (pSh). The phage clone which contained of anti-idiotypic antibody against benzo[a]pyrene, designated as A4, was chosen for further work because of highly specific to pSh. The available protein databases were searched. The A4 amino acid sequence was unique and 76% identical to a sequence in antibody against interferon . The A4 protein was expressed in bacteria and purified by two different methods: His-tagged A4 and CBD-fusion A4. Both the A4 bound to pSh and also to the human single chain idiotypic antibody against the benzo[a]pyrene (T72) by ELISA. The K values of A4 for pSh and T72 were very close: 4.44 × 10 M and 5.71 × 10M, respectively. A4 was a competitor with benzo[a]pyrene for binding sites of both idiotypic pSh and T72 in competitive ELISA. Thus, A4 was a high affinity anti-idiotypic against benzo[a]pyrene which recognised pSh and T72 active sites.
用针对苯并[a]芘（pSh）的鼠单链独特型抗体筛选来自健康人淋巴细胞的天然文库。由于对pSh具有高度特异性，选择了含有抗苯并[a]芘独特型抗体的噬菌体克隆，命名为A4，用于进一步研究。搜索了可用的蛋白质数据库。A4氨基酸序列是独特的，与抗干扰素抗体中的一个序列有76%的同一性。A4蛋白在细菌中表达，并通过两种不同方法纯化：His标签化的A4和CBD融合的A4。通过酶联免疫吸附测定（ELISA），A4既与pSh结合，也与抗苯并[a]芘的人单链独特型抗体（T72）结合。A4对pSh和T72的解离常数（K值）非常接近：分别为4.44×10⁻⁸M和5.71×10⁻⁸M。在竞争性ELISA中，A4是苯并[a]芘与独特型pSh和T72结合位点的竞争剂。因此，A4是一种高亲和力的抗苯并[a]芘独特型抗体，可识别pSh和T72的活性位点。
Zotero 插件