Premium
Purification of Human IgE and Rabbit Anti‐Human IgE
Author(s) -
LøWENSTEIN HENNING,
WEEKE BENT
Publication year - 1976
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1976.tb03837.x
Subject(s) - immunoglobulin e , cyanogen bromide , affinity chromatography , ion chromatography , sepharose , chemistry , antibody , chromatography , agarose , cyanogen , radioimmunoassay , biochemistry , immunology , biology , enzyme , peptide sequence , organic chemistry , gene
Summary. Human IgE from serum containing 60 μg ml ‐l was isolated and purified using a combination of gel chromatography, cation exchange chromatography, anion exchange chromatography, and affinity chromatography using con A‐Sepharose®. 1. Anti‐IgE was produced by immunizing rabbits with the IgE‐containing fraction. Specific anti‐IgE was obtained from the isolated rabbit immunoglobulins after absorption of the antibodies against non‐IgE serum proteins on Sepharose 4B, to which was coupled normal human serum enriched withχ and λ chains and a commercial anti‐IgE. 2. The IgE‐containing fraction was coupled to cyanogen bromide activated agarose and used for the production of immunoabsorbed rabbit anti‐human IgE, which was then labelled with 126 I for use in RAST and CRIE.