Studies on Generation of Human-human Hybridomas and Optimization of Antibody Production

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1. Human-human hybridomas producing monoclonal antibodies (MoAb) specific to human breast cancer cells were isolated and used then for detection of human breast cancer specific antigens. One of MoAb reacted specifically with about 40 k dalton antigens in human breast cancer McF-7 cells.2. We screened widely immunoglobulin production stimulating factor (IPSF) and found that culture super natants or lysates of human lung adenocarcinoma PC-8, human lymphoblastoid HO-323 and namalwa cells had IPSF activity. The IPSF activity was also found in lactoferrin, skim milk and its main component casain, chick egg yolk lipoprotein (YLP), and phosphate salts. These IPSFs except YLP stimulated IgM production of human-human and mouse-mouse hybridomas, but did not IgG production of IgG producers. YLP solely stimulated both IgM and IgG production of hybridomas. These results suggest that IgM production of hydridomas is regulated diffevently with IgG production3. Serum-free high-density culture of human-human hybridomas was studied for mass production of humon monoclonal antibodies. Hybridomas derived from NAT-30, a human parent cells isolated from Namalwa cells, proliferated well in serum-free medium up to 10^7 cells/ml and Ig productivity of the hybridomas at the high cell density was even higher than those at lower cell densities. Onthe contrary, hybridomas derived from HO-323 cells, another hunan parent cells, lost viability and IG productivity guickly at the high cell density. These results suggest that applicability to serum-free high-density culture is varies with hybridomas.