Expression of Immunoglobulin G. in the Cells of Hansenula Polymorpha

The melhylotrophic yeast cells of Hansenula polyinorph LR9, lira 3 and HM1-39, ura 3 and leu 2 were used as host strains lor the expression of the Fab fragment of MAK33 antibody. The MAK33 antibody reacts specificically wilh creatine kinase-M. Crealine kinase is an enzyme that is accumulated in the blood, brain, skeletal muscles, and in heart infarction. The eDNA of kappa or gamma chains were inserted in between FMD or MOX promoter and MOX terminator in the expression plasmids. In addition, the secretion signal sequence of the mating tacior-a ipre-pro- segment) and a fragment from glucoamylase peptide were also inserted in the expression plasmids for efficient production and serection. Such expression plasmids contain in their structures, the autonomously replication sequences of Hunsenulti (HARS) and of E. coli (ori) for auonomously replication of the plasmids uithin the Hansenula or E. coli transformed . cells. Also, they contain the ampicillin-resistance gene for E. coli transformants , selection and either Hansenula UR.-1 3 or LEU 2 genes as yeast transformants marker.