Animal Cell Technology: Basic & Applied Aspects, Vol. 4, pp321-326, 1992.


Serum-free Culture of Human-human Hybridoma secreting Monoclonal Antibodies Against Stomach Cancer.

AM. Ali, Midori Hayashida, Takuya Sugahara, Koichi Akiyama, MN. Dahan, and Hiroki Murakami


Abstract

Three human-human hybridomas (EMK-F7, FMK-H3 and KMK-41) were established as fusion products of lymphocytes from a donor alloimmunized with stomach cancer cells and H0-323 parent cell. EMK-F7 produces IgM and IgG which reacted specifically with MKN-28 and MKN-45 cells but not with other cell lines. FMK-H3 and KMK-41 produce IgM but reacted not only to the stomach cancer cells but also to other cell lines. When reacted with transfer-blotted cellular antigens, EMK-F7 distinguished a 50 kD antigen in MKN-28 and MKN-45 cells and 107 kD in NUGC-3 stomach cancer cell. Both hybridomas were unable to grow in ITES-ERDF or protein-free medium. When yolk lipoprotein was added into the medium, cells were able to multiply at the same rate as 5% FCS-ERDF. However, in protein-free medium, cells were able to multiply only when ferric citrate was increased to 150 g/ml.