Open AccessThe Use of Alginate Ester Films on Solid Supports in the Preparation of Water‐Insoluble Immunoadsorbents for Purification of Antigens
Author(s) -
KENNEDY John F.,
BARNES Junor A.,
BARKER S. Alan
Publication year - 1983
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1983.tb07519.x
Subject(s) - chromatography , elution , adsorption , chemistry , yield (engineering) , sepharose , cyanogen bromide , emulsion , biochemistry , materials science , organic chemistry , peptide sequence , metallurgy , gene , enzyme
A novel method developed for the preparation of immunoadsorbents is based on the technique of coating potential column supports with a propylene glycol alginate ester film created by emulsion polymerisation. The cross‐linking of the film with 1,2‐diaminoethane in alkaline solution resulted in a water‐permeable but water‐insoluble film which was stable at high pH. Sheep anti‐(human IgG) was subsequently coupled to the cyanogen‐bromide‐activated film and the resultant immunoadsorbents were used to purify IgG from whole human serum. The production of the film‐coated supports, chemical coupling of antibody and the chromatographic characteristics of four alginate‐film‐coated supports are described. With the use of Sepharose®– alginate‐ester–sheep‐anti‐(human IgG) immunoadsorbent the overall yield for IgG from five serial adsorption‐elution cycles was 94% of the adsorbed IgG. This immunoadsorbent proved to be of high binding capacity and allowed the elution of specific antigen in almost 100% yield and high purity with minimal non‐specific interference from other serum proteins.