Preparation and evaluation of Ricinus communis agglutinin affinity adsorbents using polymeric supports

被引:17
作者
Cartellieri, S
Helmholz, H
Niemeyer, B
机构
[1] GKSS Natl Res Ctr, Inst Coastal Res Phys & Chem Anal, D-21502 Geesthacht, Germany
[2] Univ Fed Armed Forces Hamburg, Inst Thermodynam, D-22043 Hamburg, Germany
关键词
affinity chromatography; affinity adsorbent; HPLAC; ligand immobilization; lectin; Ricinus communis agglutinin; asialofetuin;
D O I
10.1006/abio.2001.5177
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A practicable and efficient procedure for preparation of Ricinus communis agglutinin (RCA) affinity adsorbents has been developed. For immobilization of RCA two different polymer-based supports, Toyopearl and TSKgel (TosoHaas), were used. RCA has been successfully immobilized onto these supports with amounts of coupled ligand between 15 and 23 mg/g dry support and corresponding coupling yields of 69-93% (w/w). The prepared affinity adsorbents were characterized concerning their binding capacity for the glycoprotein asialofetuin (ASF) and accessibility of the ligand binding sites. The high accessibility of 80% showed that steric hindrance was negligible at the present ligand density. RCA-Toyopearl was successfully applied in affinity chromatography of glycoproteins indicating its high specificity. A long-term stability test proved no change in capacity for a period of at least 12 months. High-performance affinity chromatography (HPLAC) was carried out using RCA-TSKgel. Experimental results showed that the prepared adsorbents are suitable for selective separation of glycoproteins and oligosaccharides and therefore can be used for investigations of adsorption characteristics of glycoconjugates and for laboratory-scale preparations. (C) 2001 Academic Press.
引用
收藏
页码:66 / 75
页数:10
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