Preparation method of mouse monoclonal antibody IgG2a F (ab`) 2

Abstract

The invention relates to the technical field of antibody preparation, in particular to a preparation method of a mouse monoclonal antibody IgG2aF (ab`) 2. According to the preparation method providedby the invention, an IgG2a subtype mouse monoclonal antibody is subjected to enzyme digestion treatment by adopting Bromelain enzyme, the enzymolysis process is simple, the operation is convenient, the antibody does not need to be dialyzed into an acidic buffer solution, and change of the structure and the activity of the antibody is avoided. Besides, ProteinA is used for purifying F (ab ') 2 andFab, Superdex 200 Increase10 / 30GL is used for separating the F (ab') 2 from the Fab, Fc, the F (ab ') 2 and the Fab can be separated in two steps, tedious operation steps are avoided, the fragment recovery rate reaches 70% or above, buffer solutions used in the two steps are consistent, and the buffer solution replacement step is omitted, so that the purification time is saved.

Description

technical field [0001] The invention relates to the technical field of antibody preparation, in particular to a preparation method of mouse monoclonal antibody IgG2a F(ab')2. Background technique [0002] Fragments of mouse monoclonal antibodies are currently widely used in research fields such as clinical diagnosis and treatment. After antibody fragmentation, due to the removal of certain functional domains and changes in molecular weight, it has obvious advantages over full-length antibodies: in antigen- In the study of antibody binding, the interference of Fc-related factors can be avoided, the steric hindrance effect of full-length antibodies can be avoided, and the sensitivity is higher in solid-phase antigen detection. Some in vitro diagnostic kits have false positives. After the Fc segment of the enzyme-labeled antibody is removed, the false positives of the sample can be eliminated. Therefore, F(ab')2 has great practical application value in biological products. Pre...

AI Technical Summary

Problems solved by technology

Generally speaking, the order of sensitivity of different subtypes of mouse monoclonal antibodies to proteolytic enzymes is IgG2b>IgG3>IgG2a>IgG1, mouse IgG1 can be cut into F(ab')2 by one step enzyme, and when IgG2a is cut by enzyme, Firstly, F(ab')2 is produced, which is further cut into Fab after exposing a new site, so the products of IgG2a after digestion include F(ab')2, Fab, Fc, because F(ab')2 and Fab The isoelectric point is relatively close, and ion exchange cannot separate the two; the molecular weight of F(ab')2 and Fab is relatively close, and it is difficult to separate the two by Sephacryl S-200 gel method

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