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UBC Theses and Dissertations

Development of a general ligand for immunoaffinity partitioning in two phase aqueous polymer systems Stocks, Susan Jill


The partition of erythrocytes in a two phase aqueous polymer system composed of dextran T500, poly(ethylene glycol)8000 (PEG 8000) and buffer was studied and the effect of a combination of affinity ligands, namely rabbit IgG and PEG 1900 modified monoclonal IgG, was examined as a potential cell separation technique. Several hybridoma lines secreting mouse monoclonal IgG specific for the F[sub c] receptor of rabbit IgG were produced by the fusion of immunised mice spleen cells and mouse myeloma cells. The monoclonal IgG was modified by cyanuric chloride attachment of PEG 1900. The modified monoclonal antibody partitioned predominantly into the PEG rich upper phase of a two phase aqueous polymer system containing PEG 8000 and dextran T500. The PEG-modified monoclonal IgG was used as an affinity ligand in the two phase polymer system to specifically increase the partition of rabbit anti-NN glycophorin A IgG. The rabbit IgG was used together with the PEG-modified monoclonal IgG to increase the partition of human erythrocytes. The same system had no effect on rabbit erythrocytes. In summary, it was demonstrated that a monoclonal antibody can be modified and used to alter cell partition in two phase aqueous polymer systems in an immunologically specific manner.

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