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    HIGH-SPEED IN VITRO SCREENING METHOD
    1.
    发明申请
    HIGH-SPEED IN VITRO SCREENING METHOD 有权

    公开(公告)号：US20210079381A1

    公开(公告)日：2021-03-18

    申请号：US17025954

    申请日：2020-09-18

    Applicant: EPSILON MOLECULAR ENGINEERING INC.

    Inventor： Naoto NEMOTO ,  Toshiki MIYAJIMA ,  Yuta MATSUKAWA

    IPC: C12N15/10

    Abstract: The purpose of the present invention is to provide a method for identifying antibody CDR3 clusters using high speed in vitro screening a library selected from the group consisting of cDNA library and nucleic acid aptamer library comprising: (i) preparing a positive spherical shaped structure by binding a target molecule to a spherical shaped molecule, wherein the target molecule is immobilized on the positive spherical shaped structure, wherein the positive spherical shaped structure may contain a fluorescent label; (ii) preparing a negative spherical shaped structure, wherein the target molecule is not immobilized on the negative spherical shaped structure, wherein the negative spherical shaped structure may contain a fluorescent label; (iii) forming a positive spherical shaped conjugate or a negative spherical shaped conjugate by binding a target detecting molecule capable of binding to the target molecule to the positive spherical shaped structure or to the negative spherical structure, wherein the target detecting molecule is selected from the library having a size equal to or more than 1010 or equal to or less than 1014, wherein the target detecting molecule may contain a fluorescent label; (iv) separating the positive and the negative spherical shaped conjugates using a fluorescence cell sorter; (v) selecting the separated positive and the separated negative spherical shaped conjugates at least 1 time and then eluting the selected conjugates to obtain an eluted sample; (vi) amplifying a nucleic acid in the eluted sample using PCR to obtain PCR products; (vii) separating the PCR products using the fluorescence cell sorter; and (viii) conducting amplicon sequencing for CDR3 cluster analysis to identify the antibody CDR3 clusters.

    HIGH-SPEED IN VITRO SCREENING METHOD
    2.
    发明申请
    HIGH-SPEED IN VITRO SCREENING METHOD 审中-公开

    公开(公告)号：US20190085322A1

    公开(公告)日：2019-03-21

    申请号：US16145292

    申请日：2018-09-28

    Applicant: EPSILON MOLECULAR ENGINEERING INC.

    Inventor： Naoto NEMOTO ,  Toshiki MIYAJIMA ,  Yuta MATSUKAWA

    IPC: C12N15/10

    Abstract: The purpose of the present invention is to provide a high-speed in vitro screening method for any library selected from the group consisting of a cDNA display library and a nucleic acid aptamer library. This high-speed in vitro screening method involves: (i) a step for preparing positive spherical structures formed by immobilizing a target molecule on a spherical structure and negative spherical structures having no target molecules immobilized thereon; (ii) a step in which a target detection molecule, selected from the aforementioned library having a library size of greater than or equal to 1010, is bonded on each spherical structure to obtain spherical conjugates; (iii) a step in which the spherical conjugates are sorted into positive spherical conjugates and negative spherical conjugates with a cell sorter; (v) a step for supplying the nucleic acid on the surface of the sorted spherical conjugates for PCR; (vi) and a repetition step for repeating steps (ii) to (v) above using DNA obtained by PCR.

    ANTI-SARS-COV-2 ANTIBODY
    4.
    发明公开
    ANTI-SARS-COV-2 ANTIBODY 审中-公开

    公开(公告)号：US20240117012A1

    公开(公告)日：2024-04-11

    申请号：US18276697

    申请日：2022-02-21

    Applicant: Kao Corporation ,  Epsilon Molecular Engineering, Inc.

    Inventor： Shunsuke INAURA ,  Yuta MATSUMURA ,  Takuto TOJO ,  Atsushi NAGAMI

    IPC: C07K16/10 ,  A61P31/14 ,  G01N33/569

    CPC classification number: C07K16/1003 ,  A61P31/14 ,  G01N33/56983 ,  C07K2317/569 ,  G01N2333/165

    Abstract: Provided is an antibody that binds to the N-protein of SARS-CoV-2, and a method for utilizing the antibody. An antibody that binds to SARS-CoV-2, having one or more structural domains comprising CDRs shown in the following (a) or (b): (a) CDR1 consisting of the amino acid sequence of SEQ ID NO: 1 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence, CDR2 consisting of the amino acid sequence of SEQ ID NO: 2 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence, and CDR3 consisting of the amino acid sequence of SEQ ID NO: 3 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence; or (b) CDR1 consisting of the amino acid sequence of SEQ ID NO: 4 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence, CDR2 consisting of the amino acid sequence of SEQ ID NO: 5 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence, and CDR3 consisting of the amino acid sequence of SEQ ID NO: 6 or an amino acid sequence obtained by substituting one amino acid with another amino acid in the amino acid sequence.

    High-speed in vitro screening method
    5.
    发明授权
    High-speed in vitro screening method 有权

    公开(公告)号：US10781442B2

    公开(公告)日：2020-09-22

    申请号：US16145292

    申请日：2018-09-28

    Applicant: EPSILON MOLECULAR ENGINEERING INC.

    Inventor： Naoto Nemoto ,  Toshiki Miyajima ,  Yuta Matsukawa

    IPC: C40B30/04 ,  C12N15/10 ,  C12N15/09 ,  C12Q1/68 ,  C40B40/02 ,  C40B40/08

    Abstract: The purpose of the present invention is to provide a high-speed in vitro screening method for any library selected from the group consisting of a cDNA display library and a nucleic acid aptamer library. This high-speed in vitro screening method involves: (i) a step for preparing positive spherical structures formed by immobilizing a target molecule on a spherical structure and negative spherical structures having no target molecules immobilized thereon; (ii) a step in which a target detection molecule, selected from the aforementioned library having a library size of greater than or equal to 1010, is bonded on each spherical structure to obtain spherical conjugates; (iii) a step in which the spherical conjugates are sorted into positive spherical conjugates and negative spherical conjugates with a cell sorter; (v) a step for supplying the nucleic acid on the surface of the sorted spherical conjugates for PCR; (vi) and a repetition step for repeating steps (i) to (v) above using DNA obtained by PCR.

    High-speed in vitro screening method
    8.
    发明授权
    High-speed in vitro screening method 有权

    公开(公告)号：US11634705B2

    公开(公告)日：2023-04-25

    申请号：US17025954

    申请日：2020-09-18

    Applicant: EPSILON MOLECULAR ENGINEERING INC.

    Inventor： Naoto Nemoto ,  Toshiki Miyajima ,  Yuta Matsukawa

    IPC: C40B30/04 ,  C12N15/10

    Abstract: The purpose of the present invention is to provide a method for identifying antibody CDR3 clusters using high speed in vitro screening a library selected from the group consisting of cDNA library and nucleic acid aptamer library comprising: (i) preparing a positive spherical shaped structure by binding a target molecule to a spherical shaped molecule, wherein the target molecule is immobilized on the positive spherical shaped structure, wherein the positive spherical shaped structure may contain a fluorescent label; (ii) preparing a negative spherical shaped structure, wherein the target molecule is not immobilized on the negative spherical shaped structure, wherein the negative spherical shaped structure may contain a fluorescent label; (iii) forming a positive spherical shaped conjugate or a negative spherical shaped conjugate by binding a target detecting molecule capable of binding to the target molecule to the positive spherical shaped structure or to the negative spherical structure, wherein the target detecting molecule is selected from the library having a size equal to or more than 1010 or equal to or less than 1014, wherein the target detecting molecule may contain a fluorescent label; (iv) separating the positive and the negative spherical shaped conjugates using a fluorescence cell sorter; (v) selecting the separated positive and the separated negative spherical shaped conjugates at least 1 time and then eluting the selected conjugates to obtain an eluted sample; (vi) amplifying a nucleic acid in the eluted sample using PCR to obtain PCR products; (vii) separating the PCR products using the fluorescence cell sorter; and (viii) conducting amplicon sequencing for CDR3 cluster analysis to identify the antibody CDR3 clusters.

    NOVEL IMMUNO-PCR METHOD USING cDNA DISPLAY
    9.
    发明申请
    NOVEL IMMUNO-PCR METHOD USING cDNA DISPLAY 有权

    公开(公告)号：US20210381026A1

    公开(公告)日：2021-12-09

    申请号：US17408423

    申请日：2021-08-21

    Applicant: EPSILON MOLECULAR ENGINEERING INC. ,  Saitama University

    Inventor： Naoto Nemoto ,  Hironori Anzai ,  Takeru Suzuki ,  Shigefumi Kumachi ,  Takuya Terai

    IPC: C12Q1/6804

    Abstract: The present invention provides an improved immuno-PCR method by using cDNA display comprising the steps of: immobilizing a first antibody having a binding site to a solid phase: contacting a sample fluid to said antibody to bind a target molecule in said sample fluid; contacting said target molecule to a cDNA display being composed of a backbone being composed of a double strand and a side chain having a second antigen binding site to which the second antibody is bound; and conducting polymerase chain reaction to detect said cDNA quantitatively. According to the improved immune-PCR method of the present invention, the target molecule is screened and obtained quantitatively, because it uses cDNA display being composed of one protein/peptide and one DNA.

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