The use of detergents in velocity sedimentation of cell culture IgM.
The biosynthetically radiolabelled secreted proteins of human lymphoblastoid cell lines have been analysed by sucrose gradient velocity sedimentation. The addition of detergent to the gradient buffer dramatically improves the recovery of immunoglobulins from cell line supernatants, allowing the gradient fractions to be analysed further both serologically and biochemically. The results of these analyses show that IgM is synthesised in the 19S form and can be clearly separated from other components which include free light chains. Immunoglobulin appears to form a much larger portion of the proteins secreted by human lymphoblastoid lines than has formerly been recognised, a finding which may be of practical importance for the development and exploitation of human monoclonal antibodies.
D002250:Carbon Radioisotopes; D002460:Cell Line; D002499:Centrifugation, Density Gradient / Q000379:methods; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007075:Immunoglobulin M / Q000302:isolation & purification*; D008214:Lymphocytes / Q000276:immunology; D011865:Radioisotope Dilution Technique
D002250:Carbon Radioisotopes; D003902:Detergents; D007075:Immunoglobulin M