Juridical Foundation the Chemo-sero-therapeutic Research Institute

Kowa Company Limited

Satoshi Koyanagi

Hiroshi Mizokami

Hideo Yoshizaki

Yoshinobu Miyatsu

Keishin Sugawara

Toshihiro Maeda

Kenjiro Kawatsu

Toshio Murakami

Shinichi Furukawa

Tatsufumi Onchi

Kazuhiro Komatsu

A method for purifying a modified major mite allergen obtained by the genetic recombination technique and a purified modified major mite allergen obtained by said method for purification are provided. A method for purifying a modified major mite allergen obtained by the genetic recombination technique, which comprises the purification steps: (1) Washing and recovering inclusion bodies containing a modified major mite allergen obtained by the genetic recombination technique with MF membrane; (2) Dissolving said inclusion bodies followed by refolding; (3) Concentrating a solution containing the modified major mite allergen with simultaneous removal of low molecular weight components with ultrafiltration membrane; (4) Recovering the modified major mite allergen in non-adsorbed fractions with an anion exchanger; (5) Recovering the modified major mite allergen in adsorbed fractions with a hydrophobic gel; and (6) Recovering the modified major mite allergen in adsorbed fractions with an anion exchanger, and a modified major mite allergen with high purity obtained by said method for purification.

Method for purifying modified major mite allergen

The present invention relates to a method for purifying a calcium ion-binding protein by cation exchange chromatography. The present invention provide a method for isolating and purifying a calcium ion-binding protein in a simple and efficient manner from a liquid sample containing a calcium ion-binding protein and contaminants without any pretreatment such as addition of a chelating agent. More specifically, the present invention relates to a method for purifying a calcium ion-binding protein which comprises contacting said protein with a cation exchange carrier in the presence of calcium ions to let the said protein be adsorbed to the carrier, and after washing, eluting said protein, and to a calcium ion-binding protein having substantially no contaminants obtained by the method of the present invention.

Method of purifying calcium ion-binding protein

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Kumamoto, Japan

Satoshi Koyanagi