Method for purifying protein and glucose dehydrogenase

Abstract

A method of purifying a target protein from a solution, in which the target protein containing an electron transfer protein is dissolved, with the use of liquid chromatography. The liquid chromatography is performed by introducing the above-described protein solution into a tank filled with a packing agent, thus bonding the target protein to the packing agent, removing impurities, and then eluting the target protein from the packing agent with the use of an eluent containing a hydroxycholanoic acid salt. As an example of the above protein, glucose dehydrogenase containing a protein having an activity of dehydrogenating glucose is cited. The liquid chromatography is performed by combining hydrophobic chromatography with anion exchange chromatography.

Description

TECHNICAL FIELD

[0001] The present invention relates to a method for purifying protein by using liquid chromatography. The method is used, for example, when purifying glucose dehydrogenase which is bonded to electron transfer protein. BACKGROUND ART

[0002] Development efforts have been made in many different fields of industry for biosensors incorporating an enzyme which makes a specific reaction with a specific substrate. A representative example of such biosensors is a glucose sensor utilized mainly in the field of medical care.

[0003] The glucose sensor establishes a reaction system which includes an enzyme and an electron transfer material. When using the glucose sensor, an amperometric method for example is employed to quantitate glucose. The enzyme is provided by e.g. glucose oxidase (GOD) and glucose dehydrogenase (GDH).

[0004] GOD has high substrate specificity to glucose, high thermal stability, and is less expensive than other enzymes because it can be mass-produced indust...

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