一种从淋巴增生性疾病患者血清中纯化针对DNA的催化抗体的新方法。

A novel method for purification of catalytic antibodies toward DNA from sera of patients with lymphoproliferative diseases.

作者信息

Kozyr A V

机构信息

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia.

出版信息

Biochem Mol Biol Int. 1996 May;39(2):403-13. doi: 10.1080/15216549600201441.

DOI:10.1080/15216549600201441

PMID:8799469

Abstract

A novel highly efficient method of purification of DNA-hydrolyzing antibodies from sera of patients with lymphoproliferative diseases was developed. The method suggests purification of serum sample using Cibacron Blue 3GA, ion exchange chromatography on DEAE-Sepharose FF, PrG affinity chromatography and selection of the DNA-binding fraction on DNA-Sephacryl. The application of the method allows to obtain the stable and active polyclonal preparation with increased content of DNA-hydrolyzing antibodies characterized by specific binding constant determined using heteropolymeric oligonucleotide substrate. The unusual properties of anti-DNA catalytic antibodies in respect to their interactions with DEAE ion exchanger are discussed.

摘要

开发了一种从淋巴增生性疾病患者血清中纯化DNA水解抗体的新型高效方法。该方法建议使用Cibacron Blue 3GA纯化血清样品，在DEAE-琼脂糖凝胶FF上进行离子交换色谱，进行蛋白G亲和色谱，并在DNA-琼脂糖凝胶上选择DNA结合部分。该方法的应用能够获得稳定且有活性的多克隆制剂，其中DNA水解抗体的含量增加，其特征在于使用杂聚寡核苷酸底物测定的特异性结合常数。讨论了抗DNA催化抗体与DEAE离子交换剂相互作用的异常特性。