TY - JOUR
T1 - Production of anti-RNA antibody by hybridoma cells
T2 - Purification from mixed immunoglobulin products
AU - Eilat, Dan
AU - Laskov, Reuven
PY - 1981/7
Y1 - 1981/7
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0019436172&partnerID=8YFLogxK
U2 - 10.1016/0161-5890(81)90029-8
DO - 10.1016/0161-5890(81)90029-8
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C2 - 6170883
AN - SCOPUS:0019436172
SN - 0161-5890
VL - 18
SP - 589
EP - 595
JO - Molecular Immunology
JF - Molecular Immunology
IS - 7
ER -