Affinity chromatography agarose

EC 3.4.24.3]. Purified by using /V-ethylmaleimide to activate the enzyme, and wheat germ agglutinin-agarose affinity chromatography [Callaway et al. Biochemistry 25 4757 1986],... 

Cummings, R., and Kornfeld, S. (1982). Fractionation of Asparagin-linked Oligosaccharides by Serial Lectin-agarose Affinity Chromatography, ./ Biol. Chem. 257 11235-11240. 

Sepharose (e.g. Sepharose CL and Bio-Gel A) is a bead form of agarose gel which is useful for the fractionation of high molecular weight substances, for molecular weight determinations of large molecules (molecular weight > 5000), and for the immobilisation of enzymes, antibodies, hormones and receptors usually for affinity chromatography applications. 

TES-45 and TES-55 are two glycoproteins that have yet to be identified at a genetic level, but evidence has been obtained that they may also be lectins. Carbohydrate affinity chromatography with mannose-agarose shows that TES-32 selectively binds as expected, but that TES-45 is also present in small amounts (Loukas et al., 1999) unlike TES-32, TES-45 does not bind to A -ace Lylgalac t< isamine. No sequence information has yet been obtained on TES-45, but it is recognized by polyclonal antibodies generated to TES-32,... 

Cuatrecasas, P. (1970). Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J. Biol. Chem. 245, 3059-3065. 

Bethell, G.S., Ayers, J.S., Hancock, W.S., and Hearn, M.T.W. (1979) A novel method of activation of cross-linked agaroses with l,l -carbonyldiimidazole which gives a matrix for affinity chromatography devoid of additional charged groups./. Biol. Chem. 254, 2572-2574. 

Cuatrecasas, P., and Parikh, I. (1972) Adsorbents for affinity chromatography. Use of N-hydroxysuccin-imide esters of agarose. Biochemistry 11, 2291-2299. 

Inman, J.K. (1985) Functionalization of agarose beads via carboxymethyladon and aminoethylamide formation. In Affinity Chromatography-A Practical Approach (P.D.G. Dean, W.S. Johnson, and F.A. Middle, eds.), pp. 53-59. IRL Press, Washington, DC. 

March, S.C., Parikh, I., and Cuatrecasas, P. (1974) A simplified method for cyanogen bromide activation of agarose for affinity chromatography. Anal. Biochem. 60, 149-152. 

Figure 6.14 Schematic representation of the principle of biospecific affinity chromatography. The chosen affinity ligand is chemically attached to the support matrix (agarose bead) via a suitable spacer arm. Only those ligands in solution that exhibit biospecific affinity for the immobilized species will be retained...

Figure 3 Biosynthesis and purification of 90-kD elastin analogue analyzed by denaturing polyacrylamide gel electrophoresis (10-15% gradient, visualized by silver staining). Lanes 1-7 time course of target protein expression at 0, 30, 60, 90, 120, 150, and 180 minutes after induction. Lane 9 soluble lysate of induced E. coli expression strain BLR(DE3)pRAMl. Lanes 10-13 protein fractions obtained from immobilized metal affinity chromatography of the lysate on nickel-NTA agarose (imidazole gradient elution). Lanes 8,14 protein molecular weight standards of 50, 75, 100, and 150 kD.

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