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1. (WO2010123080) METHOD FOR PRODUCING SOLUBLE SR-A PROTEIN AND EVALUATION METHOD USING SAME
Latest bibliographic data on file with the International Bureau   

Pub. No.: WO/2010/123080 International Application No.: PCT/JP2010/057179
Publication Date: 28.10.2010 International Filing Date: 22.04.2010
IPC:
C12N 15/09 (2006.01) ,C07K 14/705 (2006.01) ,C07K 16/28 (2006.01) ,C12P 21/02 (2006.01) ,G01N 33/15 (2006.01) ,G01N 33/50 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
14
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
435
from animals; from humans
705
Receptors; Cell surface antigens; Cell surface determinants
C CHEMISTRY; METALLURGY
07
ORGANIC CHEMISTRY
K
PEPTIDES
16
Immunoglobulins, e.g. monoclonal or polyclonal antibodies
18
against material from animals or humans
28
against receptors, cell surface antigens or cell surface determinants
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
21
Preparation of peptides or proteins
02
having a known sequence of two or more amino acids, e.g. glutathione
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
15
Medicinal preparations
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
Applicants:
湯浅 浩司 YUASA Koji [JP/JP]; JP (UsOnly)
宮田 千鶴 MIYATA Chizuru [JP/JP]; JP (UsOnly)
濱本 あきば HAMAMOTO Akiba [JP/JP]; JP (UsOnly)
松川 泰治 MATSUKAWA Taiji [JP/JP]; JP (UsOnly)
松居 雄毅 MATSUI Takeki [JP/JP]; JP (UsOnly)
山田 泰正 YAMADA Yasumasa [JP/JP]; JP (UsOnly)
山田 一郎 YAMADA Ichiro [JP/JP]; JP (UsOnly)
ユーハ味覚糖株式会社 UHA Mikakuto Co., Ltd. [JP/JP]; 奈良県大和郡山市今国府町123-8 123-8, Imagocho, Yamatokooriyama-shi, Nara 6391031, JP (AllExceptUS)
Inventors:
湯浅 浩司 YUASA Koji; JP
宮田 千鶴 MIYATA Chizuru; JP
濱本 あきば HAMAMOTO Akiba; JP
松川 泰治 MATSUKAWA Taiji; JP
松居 雄毅 MATSUI Takeki; JP
山田 泰正 YAMADA Yasumasa; JP
山田 一郎 YAMADA Ichiro; JP
Agent:
柳野 隆生 YANAGINO Takao; 大阪府大阪市淀川区宮原1-15-1、ノスクマードビル Noskmard building,1-15-1,Miyahara, Yodogawa-ku, Osaka-shi, Osaka 5320003, JP
Priority Data:
2009-10400522.04.2009JP
Title (EN) METHOD FOR PRODUCING SOLUBLE SR-A PROTEIN AND EVALUATION METHOD USING SAME
(FR) PROCÉDÉ DE PRODUCTION D'UNE PROTÉINE SR-A SOLUBLE ET PROCÉDÉ D'ÉVALUATION L'UTILISANT
(JA) 可溶化SR-A蛋白質の製造方法及びそれを用いた評価方法
Abstract:
(EN) Disclosed is an evaluation system for a ligand such as a denatured LDL, which is highly useful in screening an agonist or antagonist to a soluble SR-A protein capable of forming a polymeric conformation, constructed by developing a means for obtaining said protein. Also disclosed are a vaccine preparation and an antibody preparation which are usable as arteriosclerosis inhibitors. A soluble SR-A protein capable of binding to a ligand can be produced by: producing a soluble SR-A protein by culturing cultured animal cells, into which a recombinant SR-A gene having a mutation in a base sequence encoding the amino acid sequence from the N-terminus to the transmembrane domain of SR-A protein or lacking said base sequence has been transferred, in a serum-free medium; and purifying the liquid culture medium containing the soluble SR-A protein, which has been optionally treated, to recover the soluble SR-A protein. The soluble SR-A protein has a polymeric conformation and, therefore, is usable as a vaccine preparation and an antigen for preparing a neutralizing antibody.
(FR) L'invention porte sur un système d'évaluation pour un ligand tel qu'un LDL dénaturé, qui est hautement utile dans le criblage d'un agoniste ou d'un antagoniste vis-à-vis d'une protéine SR-A soluble capable de former une conformation polymère, construite par développement d'un moyen pour obtenir ladite protéine. L'invention porte également sur une préparation de vaccin et sur une préparation d'anticorps qui sont utilisables comme inhibiteurs de l'artériosclérose. Une protéine SR-A soluble capable de se lier à un ligand peut être obtenue par : production d'une protéine SR-A soluble par culture de cellules animales cultivées dans lesquelles un gène SR-A recombinant ayant une mutation dans une séquence de bases codant pour la séquence d'acides aminés de l'extrémité N-terminale au domaine transmembranaire de la protéine SR-A ou ne présentant pas ladite séquence de bases a été transféré, dans un milieu exempt de sérum ; et purification du milieu de culture liquide contenant la protéine SR-A soluble, qui a été facultativement traitée, pour récupérer la protéine SR-A soluble. La protéine SR-A soluble a une conformation polymère et, par conséquent, est utilisable comme préparation de vaccin et comme antigène pour préparer un anticorps de neutralisation.
(JA)  多量体のコンフォメーションを形成する可溶化SR-A蛋白質を得るための手段を開発し、該蛋白質のアゴニストやアンタゴニストのスクリーニングに極めて有用な、変性LDL等のリガンドに対する評価系の構築を提供する。さらには動脈硬化抑制剤となるワクチン製剤や抗体製剤を提供する。SR-A蛋白質のN末端から膜貫通ドメインまでのアミノ酸配列をコードする塩基配列を変異又は欠失させた組換えSR-A遺伝子を導入した動物培養細胞を、無血清培地で培養して可溶化SR-A蛋白質を産生させ、可溶化SR-A蛋白質を含む培養液又はその培養処理液を精製して可溶化SR-A蛋白質を回収することで、リガンド結合能を有する可溶化SR-A蛋白質の製造が可能になった。可溶化SR-A蛋白質は、多量体のコンフォメーションを持つことにより、ワクチン製剤、中和抗体作製用抗原として利用することが可能である。
Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM)
European Patent Office (EPO) (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, ML, MR, NE, SN, TD, TG)
Publication Language: Japanese (JA)
Filing Language: Japanese (JA)