CONSTRUCTION METHOD FOR OCH1-DEFECTIVE ANTI-CD20 TETRAVALENT ANTIBODY EXPRESSION PICHIA PASTORIS

摘要

Provided is a construction method for OCH1-defective anti-CD20 tetravalent antibody expression Pichia pastoris, comprising: knocking out an OCH1 encoded α-1, 6-mannosyltransferase of a strain JC308; connecting the modified och1 with no expression ability to a pPICZαA with a screening marker; introducing a wild Pichia pastoris JC308, and after homologous recombination, screening to obtain an OCH1-defective strain, namely Δoch1; connecting a synthesized anti-CD20 tetravalent antibody sequence to a Pichia pastoris expression plasmid pPIC9, then introducing into the OCH1 gene-defective strain Δoch1 to obtain the anti-CD20 tetravalent antibody expression strain.