Novel purification method of human TNF-alpha (tumor necrosis factor- alpha) protein

Abstract

The invention relates to the technical field of biologics, and in particular to a novel purification method of a human TNF-alpha (tumor necrosis factor- alpha) protein. The method comprises the following steps: performing fine purification on a human TNF-alpha protein through an ion column and a hydrophobic chromatography column in sequence, and performing desalting treatment between two times of ion column treatment, wherein the ion column is at least one negative ion column and / or a positive ion column. The invention provides a TNF-alpha protein purification method which is convenient, rapid and capable of implementing large-scale sample treatment. The method is based on an AKTA explorer full-automatic protein purification system and is simple and convenient to operate, meanwhile automatic sample loading, automatic washing, on-line monitoring, real-time observation and the like of samples can be achieved, and the purification process can be adjusted according to sample situations and later sample purification requirements in time. Finally, the invention defines a set of precise purification method for TNF-alpha, after different steps of purification of a cation exchange column, a desalting column, an anion exchange column and a hydrophobic chromatography column, the purity of an obtained target protein is up to 95%, and the final protein purification yield is up to about 70%.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a novel purification method of human TNF-α protein. Background technique [0002] Tumor necrosis factor (TNF) is mainly produced by activated macrophages, NK cells and T lymphocytes. In 1985, Shalaby named the TNF produced by macrophages as TNF-α, and the lymphotoxin (LT) produced by T lymphocytes as TNF-β. The biological activity of TNF-α accounts for 70% to 95% of the total activity of TNF, so TNF is often referred to as TNF-α. [0003] The functions of TNF-α are mainly to kill or inhibit tumor cells, improve the phagocytic ability of neutrophils, resist viruses and parasitic infections, promote the differentiation of myeloid leukemia cells into macrophages, and promote cell proliferation and differentiation. It is a very important primer in the process of animal somatic cell immunity, and plays an important role in the process of animal anti-tumor and anti-infection. [0004] M...

AI Technical Summary

Problems solved by technology

[0004] Most existing purification methods for TNF-α have problems such as single purification method, low product purity, and excessive loss of activity. However, the multimeric form of TNF-α has more important natural activities than the monomeric TNF-α