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Selective adsorption of proteins to their ligands covalently immobilized onto microfibers
Author(s) -
Kato Koichi,
Ikada Yoshito
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260470508
Subject(s) - chemistry , carbodiimide , ligand (biochemistry) , acrylic acid , dissociation constant , adsorption , protein adsorption , covalent bond , polymer chemistry , combinatorial chemistry , chromatography , biochemistry , biophysics , organic chemistry , polymer , copolymer , receptor , biology
Following ozone oxidation of polyester microfibers of 3.5 μm average diameter and 0.83 m 2 /g specific area, the fiber surface was subjected to graft polymerization of acrylic acid and subsequently immobilized with serologically active proteins including Staphylococcus aureus protein A, a specific antigen, and a specific antibody. The immobilization reaction was mediated by a watersoluble carbodiimide, which allowed formation of a co‐valent linkage between the ligand proteins and the grafted poly(acrylic acid)chains. The yields of the immobilized ligand proteins were of the order of 1 mg/g fiber. Their binding affinity and capacity to respective specific proteins were studied in vitro from a buffered solution and serum. It was found that the specific proteins were selectively adsorbed with dissociation constants as low as 1× 10 −6 M , suggesting the adsorption to take place through highly specific protein‐protein interaction. An addition of serum albumin did not significantly affect the specific binding, regardless of the ligand proteins. The binding capacity ranged from 1 × 10 −13 to 1× 10 −11 mol/cm 2 primarily depending on the surface density of the immobilized ligands and the number of their binding sites per molecule. © 1995 John Wiley & Sons Inc.