Use of biospecific interactions of collagen, fibronectin and their fragments in affinity chromatography.
Various aspects of the application of fibronectin-collagen biospecific interactions in affinity chromatography are described. A new biospecific method for one-stage isolation of collagen peptides containing fibronectin-binding sites is proposed. The alpha 1 CB7-peptide of type-I collagen cyanogen bromide cleavage was isolated by means of affinity chromatography on adsorbents containing an immobilized gelatin-binding domain (45,000 relative molecular mass) of fibronectin. The method gives highly purified preparations of alpha 1 CB7-peptide. This peptide, as well as some other collagen molecular fragments (alpha-chains, beta-components, alpha 1 CB8-peptide), were immobilized on Sepharose, and the properties of such affinity adsorbents obtained were studied. Adsorbents with immobilized alpha-chains and alpha 1 CB7-peptide had a fibronectin-binding capacity 1.5-2.0 times higher than commercial gelatin-Sepharose. Large-scale production of highly purified fibronectin from human plasma, using affinity chromatography on immobilized individual alpha-chains of collagen, was developed.
Klyashchitsky
B A
BA
Institute of Biological and Medical Chemistry, Russian Academy of Medical Sciences, Moscow.
Mitina
V Kh
VKh
Frantsuzova
N A
NA
Krasnopolsky YuM
Shvets
V I
VI
D001665:Binding Sites; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet
D005353:Fibronectins; D010446:Peptide Fragments; D003094:Collagen