title: Two-step chromatography purification of IgGs possessing sialidase activity from human blood serum.
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.
authors.affiliation: Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov St., 14/16, 79005, Lviv, Ukraine.
authors.forename: Yury
authors.initials: Y
authors.lastname: Kit
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Bilyy
authors.forename: Nataliya
authors.initials: N
authors.lastname: Korniy
authors.forename: Andriy
authors.initials: A
authors.lastname: Tomin
authors.forename: Valentyna
authors.initials: V
authors.lastname: Chop'yak
authors.forename: Yaroslav
authors.initials: Y
authors.lastname: Tolstyak
authors.forename: Volodymyr
authors.initials: V
authors.lastname: Antonyuk
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Stoika
mesh_terms: D000818:Animals; D017151:Antibodies, Catalytic; D002417:Cattle; D002624:Chemistry, Clinical / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism*; D020945:Lupus Vasculitis, Central Nervous System / Q000097:blood; D009077:Mucins / Q000737:chemistry; D009439:Neuraminidase / Q000378:metabolism; D012685:Sepharose / Q000737:chemistry; D044967:Serum / Q000737:chemistry*; D013363:Submandibular Gland / Q000737:chemistry
chemical_list: D017151:Antibodies, Catalytic; D007074:Immunoglobulin G; D009077:Mucins; D012685:Sepharose; D009439:Neuraminidase
keywords: affinity chromatography; antibody; blood serum; mucin; protein G; sialidase activity