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SCREENING METHOD FOR SINGLE-CHAIN ANTIBODIES, AND SINGLE-CHAIN ANTIBODIES

外国特許コード F170009103
整理番号 (S2016-0105-N0)
掲載日 2017年6月14日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP082988
国際公開番号 WO 2017082214
国際出願日 平成28年11月7日(2016.11.7)
国際公開日 平成29年5月18日(2017.5.18)
優先権データ
  • 特願2015-219765 (2015.11.9) JP
  • 特願2016-024016 (2016.2.10) JP
発明の名称 (英語) SCREENING METHOD FOR SINGLE-CHAIN ANTIBODIES, AND SINGLE-CHAIN ANTIBODIES
発明の概要(英語) The present invention addresses the problem of providing: a screening method for single-chain antibodies having an extremely large capacity to bind with antigens, said method having excellent separation efficiency; and single-chain antibodies obtained using said screening method. This problem is solved by a screening method for single-chain antibodies that bind to antigens, said method comprising a step in which antigens bound to multilamellar liposomes are prepared, a step in which a phage library that displays single-chain antibodies is prepared, and a step in which phages displaying single-chain antibodies that bind to the antigens bound to the multilamellar liposomes are selected from the phage library.
従来技術、競合技術の概要(英語) BACKGROUND ART
In the form of a molecular target drug, antibody pharmaceutical, a medicament including a low molecular weight, peptide pharmaceuticals, as well as in vivo protein therapeutic such as cytokines, siRNA, aptamers, various studies, have been developed (for example, patent document 1). The use of antibodies as therapeutic agents, from its specificity, diseased cells expressing the specific antigen useful in the treatment of a disease. Antibody, a protein as an antigen expressed on the cell surface bound, the bound cells were effectively acts. Antibody, longer half-life in blood, specifically to the antigen having a feature of higher degree, is also very useful as antitumor agents.
As a method for obtaining such an antibody, an antibody library using a panning ('gold dust can wash the sand from' the 'panning' is referred to as likened to.) By known technology for acquiring antibodies. For example, the variable regions of a human single-chain antibody (scFv) by phage display method as expressed on the surface of, a method for selecting phages binding to antigen is known. Selected phage gene with respect to the horizontal, which binds to an antigen of a human antibody variable region encoding DNA sequence can be determined. DNA sequence of scFv that binds to an antigen if clear, the obtained sequence is incorporated into an appropriate expression vector, human antibodies can be easily manufactured (for example, Patent Document 2, 3).
In this way, an antibody library by panning using antibody antibody pharmaceuticals such as a candidate of the known method of acquiring. However, in the art, when coupling a phage antigen, antigen of a polystyrene tube or plate normally fixed, blocking or if operation is needed, and the like adsorbed to the tube and which may cause protein denaturation. As a result, some of the phage library, or a protein using the blocking those which bind to the denatured protein occurs, for the purpose of phage binding to an antigen on an antigen other than acquisition of a problem.
On the other hand, a method of obtaining the antibody described above on the other hand, multilamellar liposomes from peptide libraries using those peptides that bind to a particular antibody has been developed a method for obtaining (non-patent document 1). This document also discloses that, as a model, multilamellar liposomes by an immobilized anti-octapeptide (FVNQHLCK, SEQ ID NO:35) to the antibodies, by phage display method is expressed on the surface for a peptide library applied the, octapeptide (FVNQHLCK, SEQ ID NO:35) can be selectively acquired have been identified.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • KYOTO INSTITUTE OF TECHNOLOGY
  • 発明者(英語)
  • KUMADA, Yoichi
  • HASEGAWA, Yuya
  • UCHIMURA, Seiichi
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA 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
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL 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 RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
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