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Two‐step chromatography purification of IgGs possessing sialidase activity from human blood serum
Author(s) -
Kit Yury,
Bilyy Rostyslav,
Korniy Nataliya,
Tomin Andriy,
Chop'yak Valentyna,
Tolstyak Yaroslav,
Antonyuk Volodymyr,
Stoika Rostyslav
Publication year - 2015
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.3283
Subject(s) - sialidase , chemistry , ammonium sulfate precipitation , antibody , sepharose , affinity chromatography , chromatography , mucin , biochemistry , blood proteins , neuraminidase , immunology , biology , enzyme , size exclusion chromatography
Abstract Sialation of cell surface is known to be tightly connected with tumorigenicity, invasiveness, metastatic potential and clearance of aged cells, while sialation of immunoglobulin G (IgG) molecules determines their anti‐inflammatory properties. Recently, we have found for the first time IgG‐antibodies possessing sialidase‐like activity (sialylic abzyme) in blood serum of multiple myeloma and systemic lupus erythematosis patients. This abzyme was detected in a pool of IgGs purified by a typical procedure including immunoglobulin's precipitation with ammonium sulfate and following chromatography on protein G–Sepharose column. Here we describe a novel matrix for affinity purification of sialylic abzyme that is based on using bovine submandibular gland mucin conjugated to Sepharose matrix (mucin–Sepharose). This matrix preferentially binds sialidase‐like IgGs from a pool of sialidase‐active fraction of proteins precipitated with 50% ammonium sulfate from blood serum of the systemic lupus erythematosis patients. That allowed us to develop a new scheme of double‐step chromatography purification of sialidase‐like IgGs from human blood serum . Copyright © 2014 John Wiley & Sons, Ltd.