基于大规模色谱聚焦法从牛组织中分离胸腺素β4和胸腺素β9的方法。

Large-scale chromatofocusing-based method for isolating thymosin beta 4 and thymosin beta 9 from bovine tissues.

作者信息

Roboti A, Livaniou E, Evangelatos G P, Tsoupras G, Tsolas O, Ithakissios D S

机构信息

National Centre for Scientific Research Demokritos, Institute of Radioisotopes/Radiodiagnostic Products, Aghia Paraskevi Attikis, Athens, Greece.

出版信息

J Chromatogr B Biomed Appl. 1994 Dec 2;662(1):27-34. doi: 10.1016/0378-4347(94)00386-6.

DOI:10.1016/0378-4347(94)00386-6

PMID:7894690

Abstract

A large-scale method for the isolation of thymosin beta 4 (up to 120 mg) and thymosin beta 9 (up to 40 mg) from bovine lung (up to 2 kg) was developed. The isolation protocol included tissue homogenization in 0.4 M HClO4, centrifugation, solid-phase extraction through LiChroprep RP-18 material, chromatofocusing on polybuffer exchanger PBE 94-modified Sepharose and dialysis against water. The isolated products were characterized by analytical isoelectric focusing, reversed-phase HPLC, electrospray ionization mass spectrometry and amino acid analysis. The method developed is rapid and convenient, requires no expensive equipment and can be used for the isolation of thymosin beta 4 and homologous peptides from various animal tissues.

摘要

开发了一种从牛肺（最多2千克）中分离胸腺素β4（最多120毫克）和胸腺素β9（最多40毫克）的大规模方法。分离方案包括在0.4 M高氯酸中进行组织匀浆、离心、通过LiChroprep RP - 18材料进行固相萃取、在聚缓冲液交换剂PBE 94修饰的琼脂糖上进行色谱聚焦以及对水进行透析。通过分析等电聚焦、反相高效液相色谱、电喷雾电离质谱和氨基酸分析对分离出的产物进行了表征。所开发的方法快速便捷，无需昂贵设备，可用于从各种动物组织中分离胸腺素β4和同源肽。