Immunoglobulin isolation

Hutchens, T.W. Magnuson, J.S. Yip, T.T. Secretory IgA, IgG, and IgM immunoglobulins isolated simultaneously from colostral whey by selective thiophilic adsorption. J. Immunol. Methods 1990, 128 (1), 89-99. 

IgG, and IgM immunoglobulins isolated simultaneously 15. from colostral whey by selective thiophilic adsorption. J. 

Va.ria.tions in Methods. The various immunochemical methods can differ in a number of ways. For example, the analytical reagent may be cmde antisemm, monoclonal antibodies, isolated immunoglobulin fractions, etc. The conditions under which the method is mn, detection of the antigen—antibody complex, and the techniques used to increase sensitivity or specificity of the reaction all maybe varied. 

TSK-GEL SW columns have also been used to isolate heavy and light chains from immunoglobulins. Kast et al. (7) reported how these chains were desalted and isolated on a 21.5-mm i.d. X 60-cm TSK-GEL G4000SW semipreparative column using a 0.1 trifluoroacetic acid (TEA)/40% acetonitrile mobile phase. 

Kast, E., Pathmanbhan, N., Wong, J., O Connor, B., and Klein, M. (1996). Poster presented at Protein Society 1996, Preparative Isolation of Immunoglobulin Heavy and Light Chains by Size-Exclusion Chromatogtaphy . 

Another application shows the preparative purification and polishing of a therapeutic fusion protein with a humanized recombinant IgG protein. The fusion protein was expressed by the fermentation of baby hamster kidney cells. The filtered culture supernatant (155 liters) contained 2.2 g of IgG and 75.5 g of total protein. After the immunoglobulins were isolated by expanded bed adsorption and rebuffering, the IgG fraction was bound to Fractogel EMD SOj (M). This column achieved baseline separation of complete antibodies (fusion protein) from small amounts of antibodies lacking the fusion part. The resulting highly purified IgG fraction (110 ml) was diluted to 150 ml and... 

The immune globulins are contraindicated in patients with a history of allergic reactions after administration of human immunoglobulin preparations and individuals with isolated immunoglobulin A (IgA) deficiency (individuals could have an anaphylactic reaction to subsequent administration of blood products that contain IgA). 

Edelman, G.M., Gall, W.E., Waxdal, M.J., and Konigsberg, W.H. (1968) The covalent structure of a human gG-immunoglobulin. I. Isolation and characterization of the whole molecules, the polypeptide chains, and the tryptic fragments. Biochemistry 7, 1950-1958. 

Hjelm H, Hjelm K, Sjoquist J. Protein A from Staphylococcus aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins. FEBS Lett 1972 28 73-76. 

Affinity chromatography and related techniques (e.g., thiol chromatography and IMAC) are widely used for preparative isolation because they enable a single protein or class of proteins to be selectively purified from very complex mixtures. They may be occasionally used as analytical tools. For example, protein A affinity chromatography has been used for quantitative analysis of immunoglobulins in ascites fluid.45 Information about surface-accessible histidine and phosphate groups may be obtained using IMAC. 

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