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.

[Isolation and purification of biopolymers using an affinity chromatography method. IX. Preparation, properties and use of affinity adsorbents with immobilized polypeptide fragments of collagen]. / Vydelenie i ochistka biopolimerov metodom affinnoi khromatografii. IX. Poluchenie, svoistva i primenenie affinnykh adsorbentov s immobilizovannymi polipeptidnymi fragmentami kollagena.

Synthesis and properties of new affinity adsorbents with immobilized polypeptide fragments of collagen molecule (alpha-chains, beta-components, cyanogen bromide peptides) were described. Adsorbents with alpha-chains and alpha 1CB7-peptide had fibronectin binding capacity 1.5-2.0 times higher than commercial gelatin-Sepharose. Commercial production of highly purified fibronectin from human plasma using affinity chromatography on immobilized individual alpha-chains of collagen was developed.

[Isolation and purification of biopolymers using an affinity chromatography method. X. A biospecific approach to preparing collagen-like peptides containing segments binding fibronectin]. / Vydelenie i ochistka biopolimerov metodom affinnoi khromatografii. X. Biospetsificheskii podkhod k polucheniiu kollagenovykh peptidov, soderzhashchikh uchastok sviazyvaniia s fibronektinom.

A biospecific method for one-stage isolation of collagen peptides containing the fibronectin binding site is proposed. alpha 1CB7-peptide of the type I collagen cyanogen bromide cleavage was isolated by means of affinity chromatography on adsorbents containing an immobilized gelatin-binding domain (45 kDa) of fibronectin. The method allows one to obtain, in a short time, highly purified preparation of alpha 1CB7-peptide necessary for biochemical studies.