MONOCLONAL ANTIBODIES (MABS) ARE BEING USED WITH EXPLOSIVE FREQUENCY AND IN EVER-INCREASING AMOUNTS IN BIOMEDICAL RESEARCH AND MAY PLAY AN IMPORTANT ROLE IN THE FUTURE DIAGNOSIS AND TREATMENT OF HUMAN DISEASES.

MONOCLONAL ANTIBODIES (MABS) ARE BEING USED WITH EXPLOSIVE FREQUENCY AND IN EVER-INCREASING AMOUNTS IN BIOMEDICAL RESEARCH AND MAY PLAY AN IMPORTANT ROLE IN THE FUTURE DIAGNOSIS AND TREATMENT OF HUMAN DISEASES. THIS PROJECT PROPOSES: 1. TO DESIGN A SYSTEM FOR THE FORMATION OF HYBRIDOMAS SECRETING HUMAN MABS USING IN VITRO IMMUNIZATION;AND 2. TO DESIGN A METHOD TO GROW LARGE NUMBERS OF THESE HYBRIDOMAS IN VITRO. TO REDUCE THE PROBLEMS OF CONTAMINATION OF MAB PREPARATIONS BY MICROBES, ENDOTOXINS, AND FOREIGN PROTEINS, AND TO ENHANCE THE EFFICIENCY OF HYBRID FORMATION, ALL PHASES OF HYBRIDOMA GROWTH AND FORMATION WILL BE DONE IN A COMPLETELY SERUM-FREE DEFINED MEDIUM CONTAINING LESS THAN 15 MG/ML OF PROTEIN. IN VITRO IMMUNIZATION WILL BE CARRIED OUT WITH ALCOHOL-FIXED HUMAN B-LYMPHOMA CELLS AS ANTIGEN USING A TECHNIQUE THAT RESULTS IN THE SUCCESSFUL FORMATION OF MURINE BYDRIDS IN VITRO. LARGE-SCALE GROWTH IN SERUM-FREE MEDIUM OF ANTIGEN-SPECIFIC HYBRIDS WILL BE DONE USING LARGE SPINNER FLASKS. PURIFICATION OF MABS FROM THESE CULTURES WILL BE DONE BY ULTRAFILTRATION, AMMONIUM SULFATE PRECIPITATION, AND DIALYSIS, RESULTING IN PREPARATIONS OF OVER 90% PURITY AT MINIMAL COST.