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Isolation and Characterization of Free Chain of Immunoglobulin Produced by an Established Cell Line of Human Myeloma Cell Origin

I. Chain in Culture Medium¹

From the Departments of Biochemistry Research and of Surgery, Roswell Park Memorial Institute, ³ Buffalo, New York 14203

Abstract

A -type light chain of immunoglobulin produced by an established cell line (No. 8226) of human myeloma cells from patient S was isolated in an essentially pure state from spent culture medium. The purified preparation (S--medium) showed the common physical and chemical features of -type Bence Jones proteins. At neutral pH, S--medium showed a single peak (S_{20, w} = 3.6) in the ultracentrifuge and existed as a dimer, some of which reverted to the monomer in 1 M proprionic acid. Tryptic peptide maps of S--medium indicated that this chain, produced in vitro, was a complete light chain which was composed of both "constant" and "variable" portions.

Antigenic determinants of S--medium were compared with the Bence Jones protein of patient S from whom the cell line had been derived, and with other light chain preparations. In addition to the common determinants, S--medium had particular determinants which were found only in the Bence Jones protein of patient S, but not in others. This provides strong evidence that the cell line No. 8226 is the progeny of myeloma cells from patient S.

Footnotes

¹ Part of the work was presented at the 52nd Annual Meeting of the Federation of American Societies for Experimental Biology (Immunology Section), April 15–20, 1968, in Atlantic City, New Jersey. This work was supported in part by research grant AI-8472 from the National Institute of Allergy and Infectious Diseases.

² On leave of absence from the Institute for Cancer Research, Osaka University Medical School, Osaka, Japan.

³ A unit of the New York State Department of Health.