Nitto Boseki Co., Ltd.

National Institute of Agrobiological Sciences

Tatsuya Ohashi

Katsuhiro Katayama

Toshihide Miura

Fumio Nomura

Takeshi Tomonaga

Iwao Kiyokawa

Isao Kobayashi

Toshiki Tamura

Kazuyuki Sogawa

Takenori Ochiai

Hideaki Shimada

Kumiko Sasagawa

Yoshihiko Igarashi

Hideki Sezutsu

Keiro Uchino

Toshio Kanda

Hisae Arai

Noriyuki Funahashi

The present inventors produced transgenic silkworms which comprise a promoter of a DNA encoding a protein specifically expressed in the silk gland and a DNA encoding a recombinant antibody whose expression is regulated directly or indirectly by the promoter, and which secrete the recombinant antibody into the silk gland. The recombinant antibodies produced from the silk gland of the transgenic silkworms were confirmed to be active.

Antibody-producing transgenic silkworms and methods for producing the same

Silkworms which have (i) a DNA encoding a transcriptional regulator operably linked downstream of a promoter of a DNA encoding a protein specifically expressed in the silk gland and (ii) a DNA encoding TRACP5 operably linked downstream of a target promoter of the transcriptional regulator were produced. The result showed that active TRACP5b was produced from the silkworms. This means that TRACP5 produced from the silk gland of the silkworms undergoes processing in the silk gland that is similar to the processing taking place at bone resorption sites.

Method for production of TRACP5b

Using the protein chip technology, biological samples such as sera are subjected to proteome analysis. Thus, a protein which is a human fibrinogen &#x3b1;-E chain decomposition product and has a molecular weight of 5,900, a protein which is an apolipoprotein AII decomposition product and has a molecular weight of 7,800, and a protein which is an apolipoprotein AI decomposition product and has a molecular weight of 28,000, each showing an increase or a decrease with the habit of drinking, are newly found out. By detecting or quantifying these proteins, a liver disease in a subject such as one having a problem of drinking can be diagnosed at the early stage.

Marker proteins for diagnosing liver disease and method of diagnosing liver disease using the same

Using two types of antibodies, i.e., a first antibody having a higher affinity for a target substance than for a competitive substance and a second antibody having a higher affinity for the competitive substance than for the target substance, a specimen is treated with these two antibodies. Then, the competitive substance in the specimen first binds to the second antibody and thus the ratio of the target substance to the competitive substance in the specimen is enlarged. As a result, the target substance becomes liable to bind to the first antibody and, in its turn, the reactivity of the target substance is elevated compared with the case of using the first antibody alone. Thus, the target substance in the specimen can be accurately assayed while avoiding the effects of the competitive substance contained in the specimen.

Immunoassay method and kit to be used therein

Specific and accurate immunoassay of tartrate resistant acid phosphatase 5b (TRACP 5b) in a specimen may be carried out by bonding TRACP 5b in the specimen to an antibody, subjecting the TRACP 5b bonded to the antibody to enzyme reaction with a 2-halo-4-nitrophenylphosphoric acid or a salt thereof, a substrate for TRACP 5b, and then assaying the enzymatic activity of TRACP 5b.

Method of immunoassay tartrate resistant acid phosphatase 5b and kit to be used therein

Monocolonal antibodies having a higher reactivity with tartrate-resistant acid phosphatase 5b (TRACP 5b) than tartrate-resistant acid phosphatase 5a (TRACP 5a) and having a higher specificity to TRACP 5b can be obtained by cell fusion using as antigens TRACP 5b purified from human osteoclasts. By using the monoclonal antibody, TRACP 5b in a sample can be detected specifically with a high sensitivity.

Monoclonal antibody specific to tartrate-resistant acid phosphatase 5b and use thereof

Growing community of inventors

Koriyama, Japan

Tatsuya Ohashi