High pressure liquid affinity chromatography

Details High pressure liquid affinity chromatography High pressure liquid affinity chromatography

1982-04-26

1984-02-14

Adee, John

Liquid purification or separation

Processes

Liquid/liquid solvent or colloidal extraction or diffusing...

210656, 2101982, 2105021, B01D 1508

Patent

active

044315449

ABSTRACT:
A process for the high pressure liquid affinity chromatographic separation of at least one biological or related substance from a mixture in which the contact, washing and eluting phases are performed on a binding material made from a ligand, containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm, the binding material being so constructed that at least one biological or related substance is retained on the binding material during the contact and washing phases. In one preferred embodiment the ligand is a reactive dye, especially a triazinyl dye, the matrix is agarose or silica and the spacer arm is a substituted aminohexyl group. The chromatographic procedure is preferably performed at 100-3500 psi, at a flow rate of 0.5-2.0 ml/min. The choice of washing and eluting solutions depends on the material to be separated. However buffer solution to wash the column and a desorbing agent to elute the material are preferred. When the biological substance is an enzyme the desorbing agent is preferably an enzyme substrate, cofactor, inhibitor or analogue thereof. A binding material for use in the above HPLAC procedure is also provided. This is made from a ligand containing at least one of the groups anthraquinone, phthalocyanine or aromatic azo, coupled to a matrix through a spacer arm.

REFERENCES:
patent: 4016149 (1977-04-01), Travis et al.
patent: 4118316 (1973-10-01), Talley et al.
patent: 4336161 (1982-06-01), Rosewear et al
Chromatographic and Allied Methods by Mikes, John Wiley & Sons, New York, pp. 388-391 and 407-410.
Methods in Enzymology, vol. XLV by Lorand, pp. 462-466.
P.D.G. Dean et al, J. Chromatogr., 1979, 165, 301.
D.A.P. Small et al, J. Chromatogr. 1981, 216, 175.
C. R. Lowe et al, J. Chromatogr., 1981, 215, 303.

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