Seikagaku Kogyo Kabushiki Kaisha

Seikagaku Kogyo Co. Ltd.

Seikagaku Corporation

Seikagaku Kogyo Kabushiki Kaisha (Seikagaku Corporation)

Sadaaki Iwanaga

Tetsu Saito

Toshio Oda

Shun-ichiro Kawabata

Tatsushi Muta

Takanori Nakamura

Noriaki Seki

Takashi Morita

Kyosuke Miyazaki

Shunichiro Kawabata

Kenji Takahashi

Shin Nakamura

Makoto Niwa

Motonori Ohno

This invention relates to a DNA shown by SEQ ID No. 1 and an amino acid sequence coded by said DNA. This invention also relates to a DNA and an amino acid sequence of subunit a of (1.fwdarw.3)-.beta.-glucan sensitive factor derived from amebocytes of horseshoe crab and have potent affinity to (1.fwdarw.3)-.beta.-D-glucan in cell walls of fungi. Therefore, the invention is useful for the diagnosis of fungal diseases and as antimicrobial or eradicating agent of fungi in combination with an antifungal agent.

DNA encoding horseshoe crab amebocyte lysate factor G subunit A

The present invention relates to polypeptide having a primary structure of amino acid sequence shown by Sequence List Sequence No. 1 and DNA encoding for the polypeptide. The polypeptide is obtainable by following steps (1)-(3):

Polypeptides, and preparation and DNA encoding thereof

Horseshoe crab hemocyte polypeptides, and preparation and DNA encoding

Horseshoe crab amebocyte lysate factor G subunit A

The invention provides a lipopolysaccharide (LPS) binding protein isolated from horseshoe crab. The LPS binding protein is isolated by (i) extracting the hemocyte membrane fraction of horseshoe crab with a polyethylene glycol ether type nonionic surface active agent in the presence of Ca ions, (ii) combining the extract with immobilized LPS under conditions that permit the LPS binding protein to bind the immobilized LPS to produce an LPS-LPS binding protein complex, and (iii) harvesting the LPS binding protein released from the complex in the presence of a chelating agent. The isolated LPS binding protein has a molecular weight of about 27,000 daltons as determined by SDS polyacrylamide gel electrophoresis and is operative to bind a lipopolysaccharide endotoxin. Accordingly, the isolated LPS binding protein can be used for detecting endotoxin and/or removing endotoxin from an injectable medicine.

Lipopolysaccharide binding protein and process for producing the same

This invention relates to a novel polypeptide represented by the formula: ##STR1## wherein Lys represents lysine, Trp tryptophan, Cys cystine, Phe phenylalanine, Arg arginine, Val valine, Tyr tyrosine, Gly glycine, Ile isoleucine and X a hydroxyl group or an amino group), its analogue and a method for preparing the same. The polypeptide exhibits strong affinity for lipopolysaccharide, and is useful for removing endotoxin and as a therapeutic agent of bacterial infections, and a method for preparing the same.

Polypeptide having affinity for lipopolysaccharide and method for

Lipopolysaccharide-binding polypeptide

A process for determining a bacterial endotoxin, which comprises contacting an assay sample with (A) a material selected from the group consisting of an amoebocyte lysate of horseshoe crab an a pro-clotting enzyme separated from the lysate, and (B) a peptide-type substrate of the formula (R.sub.1 --Gly--Arg--R.sub.2), wherein R.sub.1 represents a member selected from the group consisting of an L-amino acid moiety whose N-terminal is protected by a protective group, a peptide moiety consisting of an L-amino acid and protected by a protective group at its N-terminal, a D-amino acid substituted L-amino acid moiety, and a D-amino acid substituted peptide moiety consisting of an L-amino acid, and is bonded to the amino group of the glycine moiety expressed by Gly through a peptide bond; and R.sub.2 represents a moiety which is bonded to the C-terminal of an L-arginine moiety expressed by Arg through an acid amide bond and/or ester bond and can be enzymatically hydrolyzed in the presence of the material (A) and the endotoxin to liberate R.sub.2 H, and/or its mineral acid salt, and detecting the resulting R.sub.2 H in which R.sub.2 is as defined above; and the above reagent used therefor.

Process for determining bacterial endotoxin and reagents used therefor

Growing community of inventors

Fukuoka, Japan

Sadaaki Iwanaga