Method for rapidly purifying and isolating anti-actin F(ab')2 and Fc fragments

Abstract

The invention provides a method for rapidly purifying and isolating anti-actin F(ab')2 and Fc fragments, which comprises a proteolysis step and a protein A magnetic bead purification step. The method provided by the invention is simple and convenient to operate, high in purification efficiency, accurate and reliable.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, in particular to a purified isolated protein antibody F(ab') 2 and Fc fragment methods. Background technique [0002] In the field of biopharmaceuticals, monoclonal antibody drugs include homologous monoclonal antibodies and recombinant monoclonal antibodies. In recombinant monoclonal antibodies, the F(ab') 2 The Fc fragment and the Fc fragment are derived from two different species, and the two fragments are recombined by genetic engineering and then cultured to produce antibody protein. In the protein antibody structure analysis, because F(ab') 2 The structure and function analysis of each fragment has important guiding significance for the judgment and analysis of the structure and function of the whole protein. For example, due to protein post-translational modification can cause F(ab') 2 The difference between the type of oligosaccharide and the type of oligosaccharide on the Fc fragment...

AI Technical Summary

Problems solved by technology

[0004] There are following shortcoming in the method in the prior art: (1) time-consuming long

(2) High experimental cost

The laboratory needs to be equipped with purification systems, ultra-high-speed centrifuges, ultrafiltration devices or solution replacement devices and other experimental instruments. Experiments use mobile phases, ultrafiltration tubes, protein A affinity chromatography fillers and other reagent consumables, which greatly increase the cost of experiments.

(3) Large consumption of samples and reagents

(4) High requirements for experimenters

(5) Recovery rate and final sample concentration are difficult to control

After purification and separation and subsequent ultrafiltration and concentration, due to the large number of sample transfers, it is very easy to reduce the recovery rate of the sample during processing, which directly leads to the difficulty of confirming the final sample concentration, which may affect the subsequent experimental analysis.

Images