Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines

✍ Scribed by G. Köhler; S. C. Howe; C. Milstein

• Book ID: 102824717
• Publisher: John Wiley and Sons
• Year: 1976
• Tongue: English
• Weight: 589 KB
• Volume: 6
• Category: Article
• ISSN: 0014-2980
• DOI: 10.1002/eji.1830060411

✦ Synopsis

Abstract

The defects in two nonsecreting variant clones of the mouse plasmacytoma MOPC 21 (P3) were studied by tissue culture methods. The variants (NSI and NSIII) do not synthesize detectable heavy chains. NSI synthesizes, but does not secrete, light chains and NSIII does not synthesize light chain.

A screening procedure was used allowing the detection of revertant cells secreting immunoglobulin. The method is based on a hemolytic plaque assay using anti‐immunoglobulin‐coated red cells. No revertants were detected among 2 × 10^7^ cells. Both variant lines were fused to another myeloma line (PI) which secretes a complete immunoglobulin and excess light chains. Analysis of the products by isoelectric focusing showed that in the hybrids there was no reactivation of synthesis of the nonexpressed chains. The defects leading to loss of synthesis cannot therefore be complemented in the hybrid lines.

The secretion of light chain in NSI, on the other hand, could be complemented in the hybrid but the light chain was only secreted as part of a new immunoglobulin hybrid molecule.