Preparation of high capacity affinity adsorbents using new hydrazino-carriers and their use for low and high performance affinity chromatography of lectins.
Two kinds of carriers with high concentrations of hydrazino groups were prepared by simple and convenient procedures. Hydrazino-carriers (I) and (II) were obtained on incubation of epoxy-activated carriers with hydrazine hydrate and adipic acid dihydrazide, respectively. Disaccharides were coupled to the hydrazino carriers through reductive amination in the presence of sodium cyanoborohydride. The reaction time was much shorter (24 h) than that in the case of the method involving amino-Sepharose 6B (800 h) [Matsumoto, I., Kitagaki, H., Akai, Y., Ito, Y., & Seno, N. (1981) Anal. Biochem. 116, 103-110]. The glycamyl-Sepharose thus obtained showed high adsorption capacities for lectins. Glycamyl-TSKgel G3000 PW obtained by the same method with TSKgel G3000 PW, which is a hydrophobic vinyl polymer matrix for high performance gel permeation liquid chromatography, could be successfully used for the high performance liquid affinity chromatography of lectins. N-Acetylglutamic acid was coupled to hydrazino-Sepharose 4B (I) in the presence of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. The adsorbent obtained was used for the affinity chromatography of Japanese horseshoe crab lectin.
Y
Y
Ito
Y
Y
Yamasaki
N
N
Seno
I
I
Matsumoto
D000818:Animals; D001894:Borohydrides / Q000494:pharmacology; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide / Q000494:pharmacology; D004852:Epoxy Compounds / Q000494:pharmacology; D006737:Horseshoe Crabs; D006834:Hydrazines / Q000494:pharmacology; D037102:Lectins / Q000302:isolation & purification*; D008024:Ligands / Q000032:analysis; D012685:Sepharose / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D013997:Time Factors
D001894:Borohydrides; D004852:Epoxy Compounds; D006834:Hydrazines; D037102:Lectins; D008024:Ligands; C029424:hydrazine; D012685:Sepharose; C009282:sodium cyanoborohydride; D003488:Cyanogen Bromide