Fig. 1. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation under nonreducing conditions of the proteins present in UIC2 and UIC2/A monoclonal antibody preparations. Lanes 1 through 4 show immunoglobulins purified on a Protein A affinity column (1.2 p,g of protein in lanes 1-3, 0.4. p,g in lane 4) after harvesting from the ascitic fluid of the UIC2 hybridoma (lane 1), tissue culture supernatant fluid of UIC2 cells grown in medium containing 10% fetal bovine serum (lane 2), and tissue culture supernatants of UIC2/A hybridoma cells grown in PFHM II protein-free media (lanes 3 and 4). Lane 5 in the figure contains Bio-Rad molecular mass markers (200, 116, 97, 66, and 45 kDa). Lanes 6 through 9 show unfractionated proteins from the tissue culture supernatant from UIC2/A cells concentrated by centrifugation using AMICON Centriprep 100 concentration units and electrophoresed at the following amounts: 50 p,g in lane 6; 25 p,g in lane 7; 12.5 p,g in lane 8; 6.25 p,g in lane 9. Lane 10 contains 4 p,g of unpurified and unconcentrated protein from the UIC2/A suspension culture supernatant.
stability of mouse MAbs at slightly basic pH, Protein A-based purification systems are more advantageous than those based on Protein G. Mouse IgG1 and IgM MAbs can be isolated using Protein L-based purification kits.
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