Process for separating and purifying recombinant human serum albumin and fusion protein thereof

Abstract

Different from the conventional method for purifying human serum albumin from human plasma, the invention provides a simple method for separating and purifying the human serum albumin from fermentation liquor or a mammal cell culture solution. By an immobilized metal affinity chromatography (IMAC) step, the method is suitable for purifying recombinant proteins at various salt concentrations, fermentation supernate at high salt concentration is allowed to be directly loaded, the total volume of loading buffer is effectively reduced and the cost is reduced. The method is stable and high-efficiency, and is suitable for laboratories, pilot scale tests and industrial production scale-up.

Description

technical field

[0001] The invention belongs to the technical field of protein separation and purification, in particular to the purification of recombinant human serum albumin. Background technique

[0002] Human serum albumin (HSA), is the protein with the most content in plasma, accounting for 40%-60% of total plasma protein, it is a very important carrier in plasma, in the environment of body fluid pH7.4, albumin is a negative ion, Each molecule can carry more than 200 negative charges. Many poorly soluble substances can be transported by binding to albumin. These substances include bilirubin, long-chain fatty acids (4-6 molecules per molecule can be combined), bile salts, prostaglandins, steroid hormones, metal ions (such as Cu 2+ 、Ni 2+ , Ca 2+ ) drugs (such as aspirin, penicillin, etc.). Another role is to maintain osmotic pressure.

[0003] The molecular structure of albumin was elucidated in 1975. It is a globular protein with 585 amino acid residues and a mol...

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