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Download Affinity Chromatography. Biospecific Sorption–The First by O. Hoffmann-Ostenhof PDF

By O. Hoffmann-Ostenhof

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IMMOBILIZED ADENINE COENZYMES IN GENERAL LIGAND AFFINITY CHROMATOGRAPHY AND THEIR USE AS ACTIVE COENZYMES Klaus Mosbach Biochemical Division, Chemical Center, University of Lund, S-220 07 Lund 7, Sweden There is considerable advantage in affinity chromatography in a versatile ligand or "general ligand11 with a broad biological affinity-binding spectrum in that a new synthetic route leading to suitable affinity matrices does not have to be devised for every putative biospecific purification. Adenine coenzymes meet this requirement and the first examples that appeared in the literature on their application involved model separations of dehydrogenases using immobilized NAD (refs.

A considerable amount of effort has been expended on synthesizing preassembled analogs of 5'-AMP, 2,,5'-ADP, and 3',5'-ADP, which constitute the corresponding "half-molecules" of NAD, NADP, and CoA, respec­ tively. The first general ligand thus tailored for immobilization and of un­ ambiguous, defined structure was N^-(6-aminohexyl)-AMP (ref. 3) (Fig. 1). In the subsequent studies the corresponding N6-(6-aminohexyl)-2',5'-ADP and N6-(6-aminohexyl)-3',5'-ADP were prepared (ref. 4 ) . e. position N^, the exocyclic nitrogen on the adenine moiety, and by the same spacer molecule.

14) R . L a m e d , Y. L e v i n & M . W i l c h e k , C o v a l e n t coupling of n u c l e o t i d e s to a g a r o s e for affinity c h r o m a t o g r a p h y , B i o c h i m . B i o p h y s . A c t a 304, 231 (1973). (15) C-Y. Lee, D . A . Lappi, B. Wermuth, J. E v e r s e & N . O . Kaplan, 8 - ( 6 - A m i n o h e x y l ) - A m i n o - A d e n i n e N u c l e o t i d e D e r i v a t i v e s for Affinity C h r o m a t o g r a p h y , A r c h s . B i o c h e m . B i o p h y s . 163, 5 6 1 , (1974).

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