Kabushiki Kaisha Toyota Chuo Kenkyusho

Fumihiko Hoshino

Osamu Asami

Yukio Yamada

Masana Hirai

Takao Imaeda

Tsutomu Kajino

Hidehiko Sugiyama

Kiyoko Sarai

Chie Idekoba

Makoto Mouri

Osamu Watanabe

Mamiko Narita

Masahito Shiozawa

Hideo Nakane

Taiji Ikawa

A method for immobilizing microobject to a solid-phase material is provided. The method comprises a step of preparing the solid-phase material having on its surface an intermediate agent that includes a first element that is capable of interacting with at least the above microobject, and a microobject immobilizing step of immobilizing the microobject to the solid-phase material in a state in which the interaction of the intermediate agent is exhibited.

Method of producing solid-phase body having immobilized microobject and the use thereof

A method for detecting or assaying one constituting member in a specific binding pair, for example, the antigen in an antigen/antibody pair, by utilizing specific binding such as binding between an antigen and an antibody, together with redox reaction for detecting a label, wherein an oxygen micro-electrode with a sensing surface area of 1 mm or less is used; and an apparatus to which the method is applicable. According to the method and by using the apparatus, redox reaction for assaying the label can be completed in such a short time as several minutes. Therefore, an inexpensive disposable apparatus for household use can be realized.

Method for detecting or assaying target substance by utilizing oxygen electrode

A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polyeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins. Further, a process which enables the polylpeptide possessing PDI activity to be efficiently produced using Humicola insolens or a transformant transformed with an expression vector containing the above-described gene is also provided.

Polypeptide possessing protein disulfide isomerase activity gene

A highly thermostable polypeptide possessing protein disulfide isomerase (PDI) activity, a gene coding for the polypeptide and a process for producing the polypeptide are provided. The polypeptide possessing PDI activity is characterized by A) having a capability of catalyzing a disulfide exchange in proteins, B) recognizing mainly ribonuclease A as a substrate, C) having a suitable active temperature of 20.degree. to 70.degree. C., D) being stable at a pH value of 6 to 9, and E) having a molecular weight of about 60,000 to 62,000. Since it has a higher thermostability and exhibits a stable activity in a wider dithiothreitol concentration range as compared with the conventional PDI, it is possible to provide a novel enzyme active protein which can be advantageously used for a refolding reaction of certain proteins. Further, a process which enables the polypeptide possessing PDI activity to be efficiently produced using Humicola insolens or a transformant transformed with an expression vector containing the above-described gene is also provided.

Polypeptide from Humicola insolens possessing protein disulfide

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Aichi, Japan

Fumihiko Hoshino