Open AccessLarge‐scale purification of choline acetyltransferase and production of highly specific antisera
Author(s) -
OSTERMANN Christel,
DICKMANN Ulrich,
MULEY Thomas,
MÄDER Michael
Publication year - 1990
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1990.tb19217.x
Subject(s) - antiserum , choline acetyltransferase , affinity chromatography , chromatography , enzyme , specific activity , chemistry , monoclonal antibody , titer , yield (engineering) , enzyme assay , choline , biochemistry , antibody , biology , acetylcholine , immunology , materials science , metallurgy , endocrinology
Choline acetyltransferase (ChAT) was purified by immunoaffinity chromatography using a covalently immobilized monoclonal antibody. In a two‐step procedure, 10 kg porcine brain yielded 750 μg active enzyme of apparent homogeneity. This amount of ChAT was purified routinely. The purification factor was 18000 and the yield of activity 4.3%. The affinity resin was stable under the experimental conditions applied and was used many times. The highly purified enzyme was subsequently employed to obtain a specific anti‐ChAT antiserum of high titer.