Derivation of specific antibody‐producing tissue culture and tumor lines by cell fusion

Abstract

Cell fusion techniques have been used to produce hybrids between myeloma cells and antibody-producing cells. The hybrid lines derived are permanently adapted to grow in tissue culture and are capable of inducing antibody-producing tumors in mice. Spleens from mice immunized against sheep red blood cells (SRBC) were fused to an 8-azaguanine-resistant clone (X63-Ag8) of MOPC 21 myeloma. Over 50% of the derived hybrid lines produce and secrete immunoglobulins different from the MOPC 21 myeloma. About 10% of the hybrid lines exhibit anti-SRBC activity. The high proportion of antibody-producing hybrids suggests that the fusion involves a restricted fraction of the spleen cell population, probably cells committed to antibody production. In order to avoid the presence of the MOPC 21 heavy chain in the specific hybrids, another myeloma cell line (NSI/1-Ag4-1) has been used. This is a nonsecreting variant of the MOPC 21 myeloma which does not express heavy chains. Three anti-SRBC (probably of the mu, gamma2b and gamma1 classes, respectively) and two anti-2,4,6-trinitrophenyl (of the mu class) antibody-producing hybrids have been repeatedly cloned. By random selection and by selection of specific clones according to their lytic activity (clone plaque selection), a number of different lines have been constructed. Such lines express different combinations of the four possible chains of each hybrid line: the myeloma gamma and K chains and the specific antibody heavy and light chains. In three cases (Sp1, Sp2 and Sp7) it is shown that only the specific H and L combination has activity and that the myeloma chains are unable to substitute for them. In most cases lines have been derived which no longer express the MOPC 21 chains but only the specific antibody chains.

Legionella pneumophilaserogroup 1 subgrouping by monoclonal antibodies – an epidemiological tool

“…Splenocytes from these mice were then fused with P3-NS1-1-Ag. 4-1 mouse myeloma cells by standard procedures (Kohler & Milstein, 1976 …”

Development and characterization of monoclonal antibodies against human aryl hydrocarbon receptor

“…Seven adult female BALB/c mice were immunized GST-hAhRs (20 μg per mouse) for 4 times by subcutaneous injection. Subsequently, according to the standard methodology (Köhler and Milstein, 1976), spleen cells from three selected immunized mice were fused with SP2/0 cells in polyethylene glycol (PEG1500, Merck, Germany), and the hybridoma cells were cultivated.…”

Hybrid myeloma cells which secrete heterodimetric IgG: A model to study the N‐linked glycan

“…obtained from a fusion of P3X63Ag8 and a murine splenocyte and selected for resistance to 5-bromouridine. Both cell lines were developed in the laboratory of Dr C. Milstein (18) and were a gift from Dr N. Cowan. These cells and their hybrids were cultured In vitro using Dulbecco's modified Eagle's medium (DMEM) (Gibco.…”