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Coupling of effector molecules to solid supports

Development of an alternative to the cyanogen bromide activation of polysaccharides

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Abstract

In view of the limited stability of the isourea bond, formed in ligand coupling to CNBractivated polysaccharides, an alternative to this current activation method has been developed. 2,4,6-Trifluoro-5-chloropyrimidine (FCP), known as a reactive group in reactive dyes, was used to activate Sepharose. Under appropriate conditions a thermally stable product with unimpaired beaded structure was obtained, which was reactive toward amines and mercaptans. Coupling with hexamethylenediamine, aniline, and ethanethiol, respectively, yielded an incorporation of 0.2-2.7, 0.9-1.7, and 1.1 mmol ligand/g dry agarose. The stability of immobilized ligands based on FCP-Sepharose between pH 4 and 8 was about 200 times higher as compared to products originating from CNBr-Sepharose; ligand leakage was only 0.5 x 10p-3%/h. The possibility of obtaining a high degree of substitution is a further advantage of the FCP activation. In addition, the FCP-activated Sepharose can be stored in the wet state at 4°C without substantial decrease in coupling capacity. The FCP analogs 2,4,5,6-tetrachloro- and 2,4,5,6-tetrafluoropyrimidine, and other polymers (cellulose, Sephadex, aminomethylpolystyrene) appeared to be applicable also.

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Authors and Affiliations

  1. Department of Organic Chemistry, Catholic University, Toernooiveld, Nijmegen, The Netherlands

    T. C. J. Gribnau, G. I. Tesser & R. J. F. Nivard

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  1. T. C. J. Gribnau
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  2. G. I. Tesser
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  3. R. J. F. Nivard
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Gribnau, T.C.J., Tesser, G.I. & Nivard, R.J.F. Coupling of effector molecules to solid supports. Journal of Solid-Phase Biochemistry 3, 1–32 (1978). https://doi.org/10.1007/BF02991408

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