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[一种获得针对重组α-突触核蛋白的高特异性多克隆抗体的合理方法]

[A rational approach to obtaining high-specific polyclonal antibodies against recombinant alpha-synuclein].

作者信息

Barinova K V, Melnikova A K, Schmalhausen E V, Muronetz V I

机构信息

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia; Sechenov First Moscow State Medical University, Institute of Molecular Medicine, Moscow, Russia.

Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia.

出版信息

Biomed Khim. 2018 Jun;64(3):276-282. doi: 10.18097/PBMC20186403276.

Abstract

The approach for the quick and efficient production ofpolyclonal antibodies tothe target antigen alpha-synuclein has been proposed. Two methods have been employed to purify specific rabbit polyclonal antibodies against recombinant human alpha-synuclein, produced by subcutaneous immunization with complete Freund's adjuvant. It was shown that purification on CNBr-activated Sepharose with immobilized alpha-synuclein resulted in antibody preparation with rabbit serum histidine-rich glycoprotein as a contaminant. Two-stage antibody purification procedure first on Sepharose with immobilized protein G, and then on alpha-synuclein immobilized column helps to avoid contamination and to obtain homogenous antibody preparation. Antibodies recognize different conformations of alpha-synuclein and can be used in a variety of immunochemical approaches, including immunocytochemistry.

摘要

已提出快速高效生产针对靶抗原α-突触核蛋白的多克隆抗体的方法。采用了两种方法来纯化针对重组人α-突触核蛋白的特异性兔多克隆抗体,该抗体通过用完全弗氏佐剂皮下免疫产生。结果表明,在固定有α-突触核蛋白的溴化氰活化琼脂糖上进行纯化,得到的抗体制剂中含有兔血清富含组氨酸糖蛋白作为污染物。两阶段抗体纯化程序,先在固定有蛋白G的琼脂糖上进行,然后在固定有α-突触核蛋白的柱上进行,有助于避免污染并获得均匀的抗体制剂。这些抗体可识别α-突触核蛋白的不同构象,可用于多种免疫化学方法,包括免疫细胞化学。

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