Production of anti-RNA antibody by hybridoma cells: Purification from mixed immunoglobulin products

Dan Eilat*, Reuven Laskov

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Fusion between spleen cells from an autoimmune NZB/NZW mouse and the Balb/c drugresistant MPC-11 myeloma resulted in the formation of a hybridoma-secreting RNA-specific IgG-3 antibody and the parental IgG-2b myeloma. Analysis of the mixed immunoglobulin assembly products made by the hybridoma cells showed efficient pairing of IgG-2b and IgG-3 heavy chains and did not show a marked preferential assembly of the homologous heavy and light chains. Partial purification of the anti-RNA antibody from the mixed assembly products was achieved by utilizing an antigen affinity column (RNA-Sepharose). The use of a heavy chain-specific affinity column (anti-IgG-2b-Sepharose) increased the purity of the desired antibody, but parental light chains were still present after this step. A complete purification of the RNA-binding protein could be achieved by papain cleavage of the total IgG fraction and binding of the resulting Fab fragments to RNA-Sepharose. This procedure may, therefore, be employed as a general method for purifying antibodies from hybridomas that continue to produce their parental myeloma chains.

Original languageEnglish
Pages (from-to)589-595
Number of pages7
JournalMolecular Immunology
Volume18
Issue number7
DOIs
StatePublished - Jul 1981
Externally publishedYes

Fingerprint

Dive into the research topics of 'Production of anti-RNA antibody by hybridoma cells: Purification from mixed immunoglobulin products'. Together they form a unique fingerprint.

Cite this