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METHOD FOR PRODUCING POLYMERIZED POLYPEPTIDE AGGREGATE AND METHOD FOR INCREASING MOLECULAR WEIGHT OF POLYMERIZED POLYPEPTIDE

Foreign code F210010278
File No. (S2019-0398-N0)
Posted date Jan 27, 2021
Country WIPO
International application number 2020JP016297
International publication number WO 2020213568
Date of international filing Apr 13, 2020
Date of international publication Oct 22, 2020
Priority data
  • P2019-079549 (Apr 18, 2019) JP
Title METHOD FOR PRODUCING POLYMERIZED POLYPEPTIDE AGGREGATE AND METHOD FOR INCREASING MOLECULAR WEIGHT OF POLYMERIZED POLYPEPTIDE
Abstract Disclosed is a method for producing a polymerized polypeptide aggregate, said method comprising a step for reacting a polypeptide, which has a tag on the N-terminal side and/or C-terminal side, with a polymerase to give a reaction liquid containing a polymerized polypeptide; a step for freezing the reaction liquid; and a step for thawing the frozen reaction liquid, wherein the polymerase recognizes the tag.
Outline of related art and contending technology BACKGROUND ART
Vaccines, which have been developed to cope with infectious diseases, are mainly attenuated live vaccines and inactivated vaccines. recently, the demand for component vaccines based on proteins produced by recombinant techniques has been increasing. Component vaccines using recombinant proteins are safer than conventional live attenuated vaccines and inactivated vaccines, and are expected to cope with infectious diseases which cannot be handled by conventional vaccines, and have low immunogenicity. One strategy for improving immunogenicity is to increase the molecular weight of the antigen.
As a method of increasing the molecular weight of the antigen, there is a method of using a chemical crosslinking agent. For example, as reported that a highly crosslinked protein polymer can be obtained by HRP enzymatic reaction by introducing Y-tag to both ends of N / C of a monomer protein, an enzyme can be used as a chemical crosslinking agent and polymerized to increase the molecular weight of the antigen.
Scope of claims (In Japanese)[請求項1]
 N末端側及びC末端側の少なくとも一方にタグを有するポリペプチドを重合化酵素と反応させて、重合化ポリペプチドを含む反応液を得る工程、
 前記反応液を凍結処理する工程、及び
 凍結処理した前記反応液を融解処理する工程
を含む、重合化ポリペプチドの集合体の製造方法であって、
 前記重合化酵素が前記タグを認識するものである、方法。

[請求項2]
 前記ポリペプチドが、N末端側及びC末端側にタグを有するポリペプチドである、請求項1に記載の方法。

[請求項3]
 前記N末端側のタグ及びC末端側のタグがペプチドタグである、請求項1又は2に記載の方法。

[請求項4]
 前記重合化酵素がフェノール酸化酵素を含み、
 前記N末端側のタグ及びC末端側のタグがそれぞれ少なくとも一つのチロシン残基を含む、
請求項3に記載の方法。

[請求項5]
 前記ポリペプチドが単量体である、請求項1~4のいずれか一項に記載の方法。

[請求項6]
 前記ポリペプチドが抗原又はその断片である、請求項1~5のいずれか一項に記載の方法。

[請求項7]
 請求項1~6のいずれか一項に記載の方法により製造された重合化ポリペプチドの集合体。

[請求項8]
 N末端側及びC末端側の少なくとも一方にタグを有するポリペプチドを重合化酵素により重合化させた重合化ポリペプチドを、凍結処理及び融解処理することで、重合化ポリペプチドを高分子量化させる方法であって、
 前記重合化酵素が前記タグを認識するものである、方法。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • KAGOSHIMA UNIVERSITY
  • KYUSHU UNIVERSITY
  • Inventor
  • MIYATA Takeshi
  • KURIHARA Koichi
  • SATO Keita
  • KAMIYA Noriho
  • MINAMIHATA Kosuke
  • KUSAKABE Takahiro
IPC(International Patent Classification)
Specified countries 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 JO JP KE KG KH 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 ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN WS 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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