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Inorganic supports coated with N ‐substituted polyacrylamides: Application to biospecific chromatography of proteins
Author(s) -
Ivanov Alexander E.,
Kozlov Leonid V.,
Shojbonov Batozhab B.,
Zubov Vitaly P.,
Antonov Vladimir K.
Publication year - 1991
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.1130050210
Subject(s) - chemistry , chromatography , acrylamide , ethanolamine , reagent , acrylate , elution , adsorption , affinity chromatography , polymer chemistry , sepharose , polyacrylamide , organic chemistry , polymer , enzyme , monomer
Wide porous glass (WPG) chemically coated with a poly‐ N ‐(2‐hydroxyethyl)acrylamide layer is proposed as a carrier of biospecific ligands in affinity chromatography. The method of WPG chemical modification includes synthesis of the γ‐aminopropyl derivative followed by chemical adsorption of poly( p ‐nitrophenyl acrylate). Ester groups of the polyacrylate‐coated WPG can be used for coupling the ligands bearing primary amino groups. Condensation of esters with ethanolamine yields a poly‐ N ‐(2‐hydroxyethyl)acrylamide‐coated support with non‐specific adsorption properties resembling those of Sepharose 4B. Human IgG immobilized on the polyacrylate support was used for isolation of the first complement component from human serum and for its separation into subcomponents C1r, C1s and C1q by a one‐step method. An unbound part of serum may be used as the R1 reagent for determining haemolytic C1 activity. The stepwise elution of C1r, C1s and C1q from the column reflects the course of C1 breakdown after its activation on immune complex formation.