Immobilization of high-affinity heparin oligosaccharides to radiofrequency plasma-modified polyethylene

✍ Scribed by Yuan, Shengmei ;Szakalas-Gratzl, Gyongyi ;Ziats, Nicholas P. ;Jacobsen, Donald W. ;Kottke-Marchant, Kandice ;Marchant, Roger E.

• Publisher: John Wiley and Sons
• Year: 1993
• Tongue: English
• Weight: 915 KB
• Volume: 27
• Category: Article
• ISSN: 0021-9304
• DOI: 10.1002/jbm.820270614

✦ Synopsis

Oligosaccharides of heparin with high affinity for antithrombin I11 (ATIII) have been immobilized onto surface-modified NHLBI Primary Reference low density polyethylene (PE). PE was modified by radiofrequency plasma polymerized ( a 5 0 nm thick) films derived from N-vinyl-2-pyrrolidone (PPNVP) or ally1 alcohol (PPAA), and coupled by chemical derivatization to either 3-aminopropyltriethoxysilane or amino-terminated poly(ethy1ene oxide). High affinity heparin oligosaccharides (HA-heparin, anti-factor Xa activity of 592 2 120 IU/mg) prepared by partial deaminative cleavage of commercial crude heparin and fractionated by agarose-ATIII affinity chromatography, were immobilized to surface-modified PE by reductive amination. The anticoagulant activity, as determined by a chromogenic assay for the inhibition of factor Xa, was estimated to be 30-70 mIU/cm2, with binding estimated to be 56-119 ng/cm2. The highest activity was obtained for the HA-heparin immobilized to PE modified by PPNVP with a PEO spacer. Visual confirmation of ATIII binding to immobilized HA-heparin was demonstrated by a goldlabeled double antibody method with imagng by SEM.