Human ubiquitin-conjugating enzyme UbcH10 monoclonal antibody hybrid tumor DY03 and monoclonal antibody
A monoclonal antibody, DY03 technology, applied in the field of cell engineering, can solve the problems of high antibody price and limited protein verification, and achieve the effect of reducing the application cost
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[0025] Example 1 Preparation and identification of the monoclonal antibody of the present invention
[0026] 1. Preparation of monoclonal antibody DY03
[0027]1) Preparation of recombinant human ubiquitin-conjugating enzyme UbcH10 antigen
[0028] The UbcH10 gene was transferred from the liver cancer cell HepG2, and the prokaryotic recombinant expression vector pET22b(+) / UbcH10 was constructed in Escherichia coli Escherichia coli Expressed in BL21, purified by Ni Sepharose affinity chromatography purification column to obtain recombinant human UbcH10 protein with a purity of more than 90%.
[0029] 2) Immunization of mice
[0030] 6-8 week old female BALB / c mice were immunized with purified recombinant antigen.
[0031] The first immunization: After mixing the recombinant antigen with an equal volume of Freund's complete adjuvant, inject BALB / c mice subcutaneously with multiple points of 50 μg / 500 μL each;
[0032] The second immunization: After 2 weeks, take the recomb...
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