Nitto Boseki Co., Ltd.

National Institute of Agrobiological Sciences

Katsuhiro Katayama

Takeshi Nagasawa

Toshihide Miura

Katsumasa Kuroiwa

Ryo Kojima

Tatsuya Ohashi

Fumio Nomura

Yoshiro Sato

Takeshi Tomonaga

Toshiki Tamura

Isao Kobayashi

Iwao Kiyokawa

Kazuyuki Sogawa

Hideaki Shimada

Takenori Ochiai

Yoshikiyo Sasagawa

Kumiko Sasagawa

Yoshihiko Igarashi

Katsuyuki Takabayashi

Hideki Sezutsu

Keiro Uchino

Kenta Noda

Shuichi Nakatsuyama

Hisae Arai

Toshio Kanda

Chie Satokawa

Katsumasa Kuriowa

Noriyuki Funahashi

Hitoshi Matsuura

Koji Endo

Akiko Takekawa

Takahiro Tomiyama

Hirotoshi Tanaka

Satoshi Inoue

Satoshi Arai

Tatsuya Baiokemikaru Kenkyusho Ohashi

Toshihide Baiokemikaru Kenkyusho Miura

Kumiko Baiokemikaru Kenkyusho Sasagawa

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

By using an antibody which is covalently bonded to a water soluble polymer as an antibody to be used in a competitive immunoagglutination assay in a homogeneous system, a protein antigen at a high concentration can be accurately assayed in an undiluted system without resorting to dilution.

Method of assaying antigen and kit to be used therein

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

A method for determination of calcium in a sample derived from a living body, by the reaction of calcium in the sample with Chlorophosphonazo-III or a compound analogous thereto in the presence of vanadate ions, followed by determination of calcium in the sample on the basis of an optical change caused by the reaction product, is excellent in various respects as follows: it is free from the problem of absorption of carbonic acid gas caused by a high pH; it is free from the problem of use of a toxic reagent containing arsenic; it makes it possible to subject a large number of samples to determination in a short time because it can be applied to an autoanalyzer; and it permits determination in a wide range because of low sample blank values.

Reagent for determination of calcium and determination method

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

A two reagent-type kit for assaying GPT by acting GPT on L-alanine and &agr;-ketoglutarate in the presence of pyridoxal phosphate, converting the resulting pyruvate into lactate with L-lactate dehydrogenase (LDH) in the presence of reduced nicotinamide adenine dinucleotide (NADH) and measuring GPT based on a decrement of NADH, the GPT assay kit comprising a first reagent and a second reagent, one of which contains pyridoxal phosphate, L-alanine, &agr;-ketoglutarate, LDH and NADH and the other reagent contains no pyridoxal phosphate but contains L-alanine. The GPT assay kit is stable over a long period of time.

Reagent for GPT assay

By making a bilirubin oxidase derived from the genus Pleurotus to act on a living body fluid sample in the presence of at least one of a cationic surfactant such as alkyltrimethylammonium salt, etc. and a nonionic surfactant such as polyoxyethylenealkyl ether, etc., only bilirubin non-.delta. fractions of the living body fluid sample can be oxidized, while keeping a bilirubin .delta. fraction remaining as unreacted. Thus, the bilirubin non-.delta. fractions and .delta. fraction of the living body fluid sample can be exactly quantitatively determined by action of the bilirubin oxidase on the sample.

Method for assaying bilirubin .delta. fractions

A method of measuring urea nitrogen on the basis of the ultraviolet absorption of urease-GLDH by a reaction rate method, which method not only permits accurate measurement of a sample containing a high concentration of urea nitrogen but also permits accurate measurement of a sample containing polyols such as mannitol without suffering the influence of the polyols, the method comprising hydrolyzing urea in a sample with urease, reacting glutamate dehydrogenase (GLDH) with ammonia formed by the hydrolysis, in the presence of .alpha.-ketoglutaric acid (.alpha.-KG) and reduced-type nicotineamide adenine dinucleotide (NADH) or reduced-type nicotineamide adenine dinucleotide phosphate (NADPH), and measuring the reduced-type nicotineamide adenine dinucleotide (NADH) or reduced-type nicotineamide adenine dinucleotide phosphate (NADPH) for a decrease rate to measure urea nitrogen derived from the urea, and characterized in that a sulfhydryl compound is co-present with the urease.

Kit for measuring urea nitrogen

Method for measuring bilirubin

According to the present invention, there are provided a monoclonal antibody to prostate-derived acid phosphatase and a method for determination of prostate-derived acid phosphatase using the monoclonal antibody. Prostate-derived acid phosphatase is boostered in animal and spleen cells isolated from the animal are fused with myeloma cells. The hybridoma capable of producing a monoclonal antibody having extremely high specificity to prostate-derived acid phosphatase is subjected to cloning. Using the monoclonal antibody produced by the hybridoma, prostate-derived acid phosphatase in a sample can be detected with extremely high sensitivity.

Monoclonal antibody to prostate-derived acid phosphatase and method for

A method of measuring the total ketone body in a sample comprising converting acetoacetic acid in the sample to 3-hydroxybutyric acid in advance and subsequently converting the whole 3-hydroxybutyric acid including 3-hydroxybutyric acid existing in the sample originally to acetoacetic acid to lead the reduction of nicotinamide adenine dinucleotide according to the action of 3-hydroxybutyrate dehydrogenase and a sample reagent thereof is disclosed. According to this invention an assay of the total ketone body in samples can be carried out conveniently, highly precisely and quickly.

Method of measuring the total ketone body and a sample reagent

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Koriyama, Japan

Katsuhiro Katayama