WO2004087914A1 - Procede de construction d'un anticorps - Google Patents

Procede de construction d'un anticorps Download PDF

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Publication number
WO2004087914A1
WO2004087914A1 PCT/JP2004/004113 JP2004004113W WO2004087914A1 WO 2004087914 A1 WO2004087914 A1 WO 2004087914A1 JP 2004004113 W JP2004004113 W JP 2004004113W WO 2004087914 A1 WO2004087914 A1 WO 2004087914A1
Authority
WO
WIPO (PCT)
Prior art keywords
antibody
gene
chain
variable region
chain variable
Prior art date
Application number
PCT/JP2004/004113
Other languages
English (en)
Japanese (ja)
Inventor
Kazuhiko Morino
Original Assignee
Institute For Antibodies Co. Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institute For Antibodies Co. Ltd. filed Critical Institute For Antibodies Co. Ltd.
Publication of WO2004087914A1 publication Critical patent/WO2004087914A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies

Definitions

  • step (c-2) a step of removing a part of the reaction solution after step (c-1), adding a primer set capable of amplifying the H chain gene thereto, and then performing a nucleic acid amplification reaction;
  • step (b) separating the labeled target cells obtained in step (a);
  • the origin of the target antibody (evening antibody) in the present invention is not particularly limited.
  • antibodies derived from various species such as mice, rats, rabbits, egrets, guinea pigs, rabbits, bush, higgins, giants, sal, chimpanzees, and humans.
  • H chain variable region (V H DJ H) gene and L chain variable region gene (V L J L) is prepared.
  • an Fv antibody molecule, scFv antibody, Fab antibody, F (ab ') 2 antibody having an ability to bind to an antigen, or a separately prepared H chain antibody Antibodies such as IgG class, IgA class, IgD class and IgE class can be obtained by using the normal region gene and the L chain constant region gene together.
  • a fusion protein expression system can be used. That is, an expression system in which a target antibody is prepared as a fusion protein with GST (glutathion S-transferase), MBP (maltose binding protein) or the like may be used.
  • a modified antibody can be obtained by binding a low-molecular compound, a labeling substance, etc. Wear.
  • the method of the present invention is applicable to all these examples.
  • it can be used for monocloning antibodies exhibiting specific properties, such as hepatitis C virus neutralizing antibodies, from human blood.
  • an antibody gene is obtained in the process. That is, an antibody gene is obtained as a result of a series of operations including a “labeling step”, a “cell separation step”, and an “antibody gene preparation step”.
  • the present inventors have further studied with attention to this point.
  • a method for analyzing a virus mutation comprising the following steps:
  • a method for preparing an autoantibody gene comprising the following steps:
  • hVK6-2 GTCCTCGCAACTGCGGCCCAGCCGGCCATGGCC GAAATTGTGCTGACTCAGTCTCCAGACTT (SEQ ID NO: 79)
  • T4 Ligase 350 U / u.I, manufactured by Takara Shuzo
  • 10 ⁇ T4 Ligase Buffer supplied with T4 DNAIigase
  • the cells were spread on an LBGA plate and cultured at 30 ° C overnight. Plasmid was extracted from 12 of the resulting transformants and subjected to agarose gel electrophoresis to confirm the nucleotide sequence of the clone containing the insert.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention se rapporte à un procédé de construction d'un anticorps permettant la préparation d'un anticorps monoclonal en un court laps de temps, et avec une grande efficacité, ledit anticorps pouvant être utilisé de manière commune. L'invention se rapporte précisément à un procédé de construction d'un anticorps comportant les étapes suivantes (a) à (d) : (a) étape consistant à amener un antigène étiqueté préparé par étiquetage d'un antigène reconnu par un anticorps cible en contact avec des cellules contenant une cellule cible produisant l'anticorps ci-dessus de manière à lier l'anticorps étiqueté à la cellule cible ; (b) étape de séparation de la cellule cible étiquetée obtenue au cours de l'étape (a) ; (c) étape d'utilisation de la cellule cible étiquetée ainsi séparée afin de préparer un antigène porté par ladite cellule et (d) étape d'expression de l'antigène ainsi préparé au moyen d'un vecteur d'expression.
PCT/JP2004/004113 2003-03-31 2004-03-24 Procede de construction d'un anticorps WO2004087914A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2003096505A JP2006180708A (ja) 2003-03-31 2003-03-31 抗体作製方法
JP2003-096505 2003-03-31

Publications (1)

Publication Number Publication Date
WO2004087914A1 true WO2004087914A1 (fr) 2004-10-14

Family

ID=33127482

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2004/004113 WO2004087914A1 (fr) 2003-03-31 2004-03-24 Procede de construction d'un anticorps

Country Status (2)

Country Link
JP (1) JP2006180708A (fr)
WO (1) WO2004087914A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009147864A1 (fr) * 2008-06-04 2009-12-10 静岡県 Procédé d'analyse/identification d'un gène d'anticorps à un niveau unicellulaire
JP2010535474A (ja) * 2007-08-03 2010-11-25 エムユーエスシー ファウンデーション フォー リサーチ ディベロップメント ヒトモノクローナル抗体およびそれを産生する方法

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100248971A1 (en) 2007-12-03 2010-09-30 Takashi Inagaki Process for antibody production
CA2947529C (fr) 2014-05-02 2022-06-21 Shinshu University Systeme d'expression-secretion de gene d'anticorps

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993018068A1 (fr) * 1992-03-09 1993-09-16 Tanox Biosystems, Inc. Procedes permettant de selectionner des lymphocytes b peu communs specifiques d'un antigene

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993018068A1 (fr) * 1992-03-09 1993-09-16 Tanox Biosystems, Inc. Procedes permettant de selectionner des lymphocytes b peu communs specifiques d'un antigene

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
WEITKAMP J.H. ET AL: "Generation of recombinant human monoclonal antibodies to rotavirus from single antigen-specific B cells selected with fluorescent virus-like particles", J IMMUNOL METHODS, vol. 275, no. 1-2, 1 April 2003 (2003-04-01), pages 223 - 237, XP004416763 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010535474A (ja) * 2007-08-03 2010-11-25 エムユーエスシー ファウンデーション フォー リサーチ ディベロップメント ヒトモノクローナル抗体およびそれを産生する方法
US20110318758A1 (en) * 2007-08-03 2011-12-29 Musc Foundation For Research Development Human monoclonal antibodies and methods for producing the same
US8715743B2 (en) * 2007-08-03 2014-05-06 Musc Foundation For Research Development Human monoclonal antibodies and methods for producing the same
JP2015171367A (ja) * 2007-08-03 2015-10-01 エムユーエスシー ファウンデーション フォー リサーチ ディベロップメント ヒトモノクローナル抗体およびそれを産生する方法
US9709574B2 (en) 2007-08-03 2017-07-18 Musc Foundation For Research Development Human monoclonal antibodies and methods for producing the same
WO2009147864A1 (fr) * 2008-06-04 2009-12-10 静岡県 Procédé d'analyse/identification d'un gène d'anticorps à un niveau unicellulaire
JPWO2009147864A1 (ja) * 2008-06-04 2011-10-27 静岡県 １細胞レベルでの抗体遺伝子の解析・同定方法

Also Published As

Publication number Publication date
JP2006180708A (ja) 2006-07-13

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