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一种针对苯并[a]芘的新型单链抗独特型抗体的分离、制备及特性研究

Isolation, production, and characterization of a new single chain anti-idiotypic antibody against benzo[a]pyrene.

作者信息

Arnst Kristina, Studennikov Artem, Ustinov Valentin, Glushkov Andrey

机构信息

a Federal State Scientific Institute, Federal Research Centre Coal and Coal Chemistry, Siberian Branch of the Russian Academy of Sciences , Institute of Human Ecology , Kemerovo , Russia.

b Federal State Educational Institute of Higher Professional Education , Kemerovo State University , Kemerovo , Russia.

出版信息

J Immunoassay Immunochem. 2017;38(6):652-662. doi: 10.1080/15321819.2017.1384390. Epub 2017 Nov 20.

Abstract

UNLABELLED

Polycyclic aromatic hydrocarbons are chemical carcinogens which could induce the development of human cancers. Anti-idiotypic antibodies against benzo[a]pyrene (BP) are perspective for human cancer immunoprophylaxis and tumor immunodiagnostic techniques. The purpose of this study was to isolate anti-idiotypic antibodies against BP from human lymphocytes naïve phage library. The anti-idiotypic antibody, named B5, was selected. Analysis of the nucleotide and amino acid sequences B5 showed no similarity to known protein databases antibodies. B5 bound idiotypic antibodies against BP in direct and competitive ELISA. It was suggested that the B5 carried an immunological image of BP and bound the idiotypic antibodies against BP.

ABBREVIATIONS

scFv: single-chain variable fragment; Ab1: idiotypic antibodies; Ab2: anti-idiotypic antibodies; CBD: cellulose binding domain; BSA: bovine serum albumin; PBS: phosphate buffer; BP-BSA: benzo[a]pyrene-BSA conjugate; Cr-BSA: chrysene-BSA conjugate; Py-BSA: pyrene-BSA conjugate; Ac-BSA: anthracene-BSA conjugate; Ba-BSA: benz[a]anthracene-BSA conjugate; PAH: polycyclic aromatic hydrocarbons; pSh: mouse idiotypic single-chain variable fragment against benzo[a]pyrene; T72: human idiotypic single-chain variable fragment against benzo[a]pyrene.

摘要

未标记

多环芳烃是化学致癌物,可诱发人类癌症。抗苯并[a]芘(BP)的抗独特型抗体在人类癌症免疫预防和肿瘤免疫诊断技术方面具有前景。本研究的目的是从人淋巴细胞天然噬菌体文库中分离抗BP的抗独特型抗体。筛选出了名为B5的抗独特型抗体。对B5的核苷酸和氨基酸序列分析表明,它与已知蛋白质数据库中的抗体没有相似性。在直接和竞争性酶联免疫吸附测定中,B5与抗BP的独特型抗体结合。提示B5携带BP的免疫影像并能与抗BP的独特型抗体结合。

缩写

scFv:单链可变片段;Ab1:独特型抗体;Ab2:抗独特型抗体;CBD:纤维素结合结构域;BSA:牛血清白蛋白;PBS:磷酸盐缓冲液;BP-BSA:苯并[a]芘-牛血清白蛋白偶联物;Cr-BSA:屈-牛血清白蛋白偶联物;Py-BSA:芘-牛血清白蛋白偶联物;Ac-BSA:蒽-牛血清白蛋白偶联物;Ba-BSA:苯并[a]蒽-牛血清白蛋白偶联物;PAH:多环芳烃;pSh:抗苯并[a]芘的小鼠独特型单链可变片段;T72:抗苯并[a]芘的人独特型单链可变片段。

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