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METHOD FOR MANUFACTURING SUBSTANTIALLY 1FL BODIES SHAPED AS CAPSULES, AND DEVICE USED FOR SAME

Foreign code F190009862
File No. (S2018-0047-N0)
Posted date Jul 26, 2019
Country WIPO
International application number 2018JP038900
International publication number WO 2019078311
Date of international filing Oct 18, 2018
Date of international publication Apr 25, 2019
Priority data
  • P2017-202950 (Oct 19, 2017) JP
Title METHOD FOR MANUFACTURING SUBSTANTIALLY 1FL BODIES SHAPED AS CAPSULES, AND DEVICE USED FOR SAME
Abstract The present invention pertains to an electrospray device that includes a microhole array chip 10, two electrodes 20, 21 provided at a distance from each other so as to face each of the main surfaces of the microhole array chip, and a power supply 30 for applying a voltage between the two electrodes. The present invention pertains to a method for manufacturing bodies shaped as capsules, said method including: filling a space 1 in the device with a liquid being encapsulated; filling a space 2 with a liquid that is poorly soluble in the liquid being encapsulated; applying a pulse wave voltage between two electrodes; causing the liquid being encapsulated to pass through microholes in the microhole array chip; and forming bodies shaped as capsules in the liquid that is poorly soluble in the liquid being encapsulated in the space 2. The present invention is a method for conducting a μL-scale cell-free reaction in capsules in fL-scale IVC, and provides a means whereby fL-scale IVC is conduced in capsules in a manner that is simpler and more efficient than in prior-art methods.
Outline of related art and contending technology BACKGROUND ART
IVC in synthetic biology (in vitro compartmentalization( compartmentalization) ) using a technique of artificial cell bottom-up structure, the efficiency of the cell-free system to a review of the looms. Cell-free reaction production rate, and inversely proportional to the scale size of the reaction has been considered (Non-Patent Document 1). This is mainly due to, not limited to the size of the reaction space of the bulk (such as from in the microliter range) is, the non-reactive components that occurs in the noise due to the Poisson distribution. Is partitioned into 2 first, the internal activity from the external environment of the chemical environment of the interface cause separation phenomenon, causing an increase in reduction in the size of the compartment. Therefore, the limited space in the cell-free reaction (femtoliter scale) of the IVC method can be performed, in synthetic biology possibility for a stimulation.
3 IVC is generated by using two major approaches. 1) Stirring and the homogenate was the water droplets and oil-in-use (non-patent document 2) bulk emulsion. Simple, resulting in a considerable amount of poly dispersion IVC (nL scale pL from the range of the size), so that the yield cannot be predicted.
2) Micro-fluid-based system (non-patent document 3) is, for generating a single IVC can be distributed, to the generation of smaller sub-pL IVC is not limited to, the production efficiency.
3) (Patent Document 1) was determined to be a single discrete fL generated IVC can be very small. However, the frequency of a single electrospray nozzle 1kHz is used, about 5 million/sec can be generated by the droplet, thus in this system, the encapsulation reaction μl fL IVC is to scale, several hours to several days would be required. For example, 10 μl to 1fl of the IVC of the reaction of the free cell is encapsulated 55 will be time consuming.
Patent Document 1: Japanese Patent Laid-Open Publication 2017-1018
Non-Patent Document 1: Acs. Synth. Bio.2014: 3,347 Non-Patent Document 2: Non-Patent Document 3 Biomacromolecules 2005, 6, 1824-1828.: Anal. Chem. 2008, 80, 3522-3529. Patent Document 1 and Non-Patent Document 1-3 all of the description, the entirety of the specifically disclosed herein.
Scope of claims (In Japanese)[請求項1]
マイクロホールアレイチップ、
前記マイクロホールアレイチップの各主表面に対向し、間隔を開けてそれぞれ設けた2つの電極、
前記2つの電極の間に電圧を印加するための電源
を含む
エレクトロスプレー用デバイス。

[請求項2]
前記マイクロホールアレイチップのマイクロホールはチップ基板を貫通する貫通孔であり、貫通孔の両方の開口の平面形状は略円形であり、一方の開口の直径は他方の開口の直径と同一又は異なる、請求項1に記載のデバイス。

[請求項3]
前記2つの電極は、前記マイクロホールアレイチップの少なくともマイクロホールが存在する表面前面に設けられている、請求項1又は2に記載のデバイス。

[請求項4]
前記マイクロホールアレイチップの少なくとも一方の主表面に導電性薄膜層をさらに有し、導電性薄膜層はマイクロホールの各開口に対応する位置にマイクロホールの開口と略同一形状および寸法の開口を有する、請求項1~3のいずれかに記載のデバイス。

[請求項5]
前記マイクロホールアレイチップの一方の主表面と一方の電極(以下、電極A)との間隔に形成された空間1、及び前記マイクロホールアレイチップの他方の主表面と他方の電極(以下、電極B)との間隔に形成された空間2は、それぞれ液体保持機能を有する、請求項1~4のいずれかに記載のデバイス。

[請求項6]
前記空間1及び2は、密閉系であるか、あるいはそれぞれ液体の流入出口を有する流通系である、請求項5に記載のデバイス。

[請求項7]
マイクロホールの電極A側の開口の直径が10~100μmの範囲であり、マイクロホールの電極B側の開口の直径が1~10μmの範囲である、請求項1~6のいずれかに記載のデバイス。

[請求項8]
前記電源は、10~10000ボルトの範囲の電圧のパルス波を供給できる電源である、請求項1~7のいずれかに記載のデバイス。

[請求項9]
請求項5~8のいずれかに記載のデバイスの空間1に被カプセル化液を満たし、かつ空間2に被カプセル化液難溶性液を満たし、
2つの電極の間にパルス波電圧を印加して、前記被カプセル化液をマイクロホールアレイチップのマイクロホールを通過させ、空間2内の被カプセル化液難溶性液にカプセル化体を生成させることを含む、カプセル化体の製造方法。

[請求項10]
印加するパルス波電圧は、電圧が100~10000Vの範囲であり、周波数が、10~100kHzの範囲であり、デューティー比が10~90%の範囲である、請求項9に記載の製造方法。

[請求項11]
カプセル化体の平均体積が0.1fL~10fLである、請求項9または10に記載の製造方法。

[請求項12]
被カプセル化液がセルフリー反応(体)である請求項9~11のいずれかに記載の製造方法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • JAPAN ADVANCED INSTITUTE OF SCIENCE AND TECHNOLOGY
  • Inventor
  • BIYANI Manish
  • TAKAMURA Yuzuru
  • SHARMA Kirti
  • PHAN Tue Trong
  • MINAMI Noritaka
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 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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