Efficient selection of mu m-mutants from mu m-expressing myeloma cells by treatment with ricin A-conjugated anti-mu antibody.
We have developed an efficient system for obtaining myeloma mutants defective in trans-acting factors required for immunoglobulin (Ig) gene expression. The system consists of a myeloma cell line designed for this purpose and an efficient method for selecting mutants from it. The cell line is X63.653 transfected with the mu gene, whose tailpiece sequence was replaced with the transmembrane sequence of human EGF receptor to hold mu on the cell surface and whose CH1 sequence was removed to prevent mu from being retained in the endoplasmic reticulum. It efficiently and stably expressed mu chains of IgM on the cell surface (mu m+) without light chains. To obtain mutants lacking mu m (mu m-) from the mu m+ cell line by selectively killing mu m+ cells, a method with ricin A-conjugated anti-mu antibody was more reliable than complement lysis mediated by anti-mu antibody. Applying the system, we obtained a variety of mu m- mutants.
Shindo
T
T
Engineering Research Laboratories, Research Institute, Kaneka Corporation, Hyogo, Japan.
Ueda
H
H
Makishima
F
F
Suzuki
E
E
Nishimura
H
H
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D066246:ErbB Receptors / Q000235:genetics; D007075:Immunoglobulin M / Q000235:genetics*; D000922:Immunotoxins; D051379:Mice; D009101:Multiple Myeloma; D009154:Mutation; D010957:Plasmids; D012276:Ricin; D015534:Trans-Activators / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured
D007075:Immunoglobulin M; D000922:Immunotoxins; D015534:Trans-Activators; D012276:Ricin; D066246:ErbB Receptors