Date of Award

Spring 1976

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Clinical Chemistry

Committee Director

James H. Yuan

Committee Member

James Geyer

Call Number for Print

Special Collections LD4331.C45P69

Abstract

A method is described for preparing an enriched fraction of the three major antibodies, immunoglobulins IgG, IgA and IgM, from normal human serum by a batch method. Selective precipitation of these proteins was carried out in successive steps using polyethylene glycol (PEG) of average molecular weight 6,000 at concentrations of 7%, 12% and 14% (16) . Zinc sulfate (10 mM) was used to remove highly immunogenic alpha2 macroglobulin from the IgM fraction. IgA, IgM and IgG were further purified by adsorption methods using DEAE-cellulose. Quantitation and qualitation of the enriched fractions showed nearly complete separation of each of the three immunoglobulins; however, with low purity and percent, yield.

Additional experimentation using greater initial serum dilutions for the PEG precipitation, greatly enhanced yield and purity. A final non-batch method, gel filtration on a Sephadex G-200 column, resulted in an enriched fraction homogenous for each isotype immunoglobulin when assayed by immunoelectrophoresis and radial immunodiffusion.

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DOI

10.25777/w4e7-2j55

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