Use of Immunoadsorption Techniques in the Preparation of Chemical Agammaglobulinemia

Abstract

Currently the use of affinity chromatography has been directed toward selective purification of proteins (1). Specific antibodies have been obtained in high yield using columns of Sepharose coupled either to haptens or to protein antigens (2). It is the purpose of this communication to report the application of these newly developed immunoadsorbents to selective removal of immunoglobulins from serum by passage on a Sepharose anti-light chain antibody column. Preparation of the immunoadsorbent. Antiserum to guinea pig light chain was prepared by immunizing a goat with an IgG light chain pool, emulsified in complete Freund's adjuvant (3). In double diffusion analysis this antiserum reacted strongly with the κ type light chains whereas the precipitin reaction against the λ type of light chains though present was significantly weaker. Five and six-tenths grams of CNBr (37.5 mg/ml of Sepharose), dissolved in water, were added with stirring at room temperature to 150 ml of packed Sepharose 4 B (Pharmacia Chemicals, Piscataway, N. J.), contained in 300 ml of 0.05 M sodium bicarbonate buffer, pH 9.0.