PRODUCTION OF FUNCTIONAL MONOCLONAL ANTIBODIES IN YEAST

GENETIC ENGINEERING TECHNIQUES WILL BE USED TO ENABLE YEAST CELLS TO PRODUCE FUNCTIONAL MONOCLONAL ANTIBODY FRAGMENTS. THE LONG-TERM APPLICATIONS ARE: (1) RELIABLE PRODUCTION OF KILOGRAM QUANTITIES OF MONOCLONAL ANTIBODIES (NEEDED IN CANCER THERAPY AND DIAGNOSTIC IMAGING) FROM YEAST FERMENTATIONS RATHER THAN FROM MORE DIFFICULT HYBRIDOMA CULTURES, AND (2) RAPID ENGINEERING AND TESTING OF IMPROVED ANTIBODY-RELATED MOLECULES THROUGH IN VITRO RECOMBINATION AND MUTAGENESIS. FOR EXAMPLE, VECTORS CONTAINING THE HYPERVARIABLE CODING REGIONS FROM RODENT GENES CAN BE RECOMBINED WITH HUMAN CONSTANT OR FRAMEWORK-CODING REGIONS OR OTHER INTERESTING PROTEIN-CODING REGIONS TO PRODUCE NEW MOLECULES. ALSO, THE VARIABLE CODING REGIONS CAN BE RANDOMLY MUTAGENIZED IN VITRO AND RAPIDLY EXPRESSED IN YEAST CELLS. MILLIONS OF YEAST COLONIES, EACH PRODUCING A DIFFERENT SINGLY MUTATED MOLECULE, CAN BE SCREENED FOR IMPROVED ANTIBODY-BINDING SPECIFICITY AND ENZYMATIC ACTIVITY ON PETRI DISHES OR FILTERS. THE KEY TO THE SUCCESS OF THIS PROJECT IS THE RECENTLY IMPROVED TECHNOLOGY FOR SECRETION OF FOREIGN PROTEINS FROM YEAST. YEAST HOST STRAINS CAPABLE OF SECRETING SINGLE-CHAIN FV MOLECULES HAVE BEEN ENGINEERED. THEY ARE BEING TESTED FOR FAB AND INTACT ANTIBODY PRODUCTION.