High-Performance Affinity Adsorbents for Trypsin-Family Enzymes Prepared with TSKgel G3000PW(Biological)
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概要
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For high-performance liquid affinity chromatography (HPLAC) of trypsin-family enzymes, two kinds of adsorbents (I and II) were prepared by convenient procedures using m-aminobenzamidine (ABA) and TSKgel G3000PW, a carrier for high-performance gel chromatography. For adsorbent I, ABA was immobilized on succinyl-TSKgel G3000PW, which was prepared by successive epoxy-activation, amination and succinylation of TSKgel G3000PW with the aid of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (about 16μmol ABA/g wet gel, about 80% yield). For adsorbent II, ABA was immobilized on formyl-carrier, which was prepared by successive epoxy-activation, hydrolysis and periodate-oxidation of TSKgel G3000PW through reductive amination with sodium cyanoborohydride (19μmol ABA/g wet gel, 98% yield). In the case of HPLAC on adsorbent I, bovine and Streptomyces griseus trypsins were eluted at 12 and 19min, respectively, whereas the enzymes were retained on adsorbent II and were recovered efficiently with eluting buffer containing 6-aminohexanoic acid. The results indicate that adsorbent I is useful for rapid analysis of the enzymes and adsorbent II for the preparation of the enzymes.
- 1987-09-25
著者
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Matsumoto Isamu
The Graduate School Of Humanities And Sciences Ochanomizu University
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MATSUMOTO Isamu
Department of Obstetrics and Gynecology, Faculty of Medicine, Kagoshima University
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Seno N
Department Of Chemistry Ochanomizu University
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Seno Nobuko
Department Of Chemistry Faculty Of Science Ochanomizu University
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Matsumoto Isamu
Department Of Chemistry Faculty Of Science Ochanomizu University
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KANAMORI AKIKO
Department of Chemistry, Faculty of Science, Ochanomizu University
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Kanamori Akiko
Department Of Chemistry Faculty Of Science Ochanomizu University
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MATSUMOTO ISAMU
Department of Biochemistry, Kurume University School of Medicine
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