Read e-book online Affinity Chromatography PDF

By Jaroslava Turková (Eds.)

ISBN-10: 0444416056

ISBN-13: 9780444416056

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From the agreement of the KL and KI values it can soluble antilysine K I = 8 9 be concluded that the formation of the complex between chymotrypsin and antilysine is not affected by its covalent bond with Spheron. The dissociation constants for the complex of chymotrypsin and antilysine, either in solution or bound to a solid matrix, are in good agreement. 8 - 1 Od6 M). 2-10-5 M, which agrees closely with the published value of 3 lo-’ M (Walsh and Wilcox). 2 DETERMINATION OF DISSOCIATION CONSTANTS BY FRONTAL ANALYSIS The derivation of quantitative affinity chromatography by frontal analysis (Kasai and Ishii) is based on the assumption that if the total length of the agarose column is 1 and the total amount of the immobilized ligand in the adsorbent is Lt, then Lo = Lt/l, where [Lo] is the density of affinity ligands per unit length of the agarose column.

Limited the analysis t o short oligomers and were * For list of symbols see p. 33. THEORY GF AFFINITY CHROMATOGRAPHY 28 able to assume an all-or-none binding of the oligomer on to polyuridylic acid. 32) x s N F p N { [ u p ( 2-p)/(l -p)'] + 1 ) + N - 1 From eqn. 33) ,'. i' . ,','. ' .. -.. I . 0 1Qo [FREE O L l G O M E R l (MOLES/LITER 10'1 Fig. 8. 033 M phosphate buffer, 1 M NaCl, 1 mM MgCl,. 9 lW4 M. Broken line, theoretical curve. The ordinate indicates the helical content of polyuridylic acid in 1 : 2 stoichiometry of adenylic acid : uridylic acid.

Acta, 354 (1974) 121-129. , Biochem. Biophys. Res. ,38 (1970) 875-881. J. , Eur. J. , 41 (1974a) 341-345. J. , Eur. J. Biochem.. 42 (1974b) 1-6. H. ,Biopolymers, 1 (1963) 133-143. , Bailon, P. , Advan. Exp. Med. , 42 (1974) 33-42. , Tanabe, S. , Biopolymers, 14 (1975) 33-49. , 34 (1974) 13-30. Porath, J. , in H. L. 95-178. H. , Chromarographia, 4 (1971) 578-581. , Cuatrecasas, P. , J. Bwl. , 246 (1971) 196-200. , Coupek, J. ,Eiochim. Biophys, Acta, 427 (1976) 586-593. 35 Chapter 4 Application of affinity chromatography to the quantitative evaluation of specific complexes Association processes undoubtedly play one of the most important roles in biochemical processes.

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Affinity Chromatography by Jaroslava Turková (Eds.)

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