Processing

Please wait...

Settings

Settings

Goto Application

1. WO2020135546 - METHOD FOR HIGH-THROUGHPUT CONSTRUCTION OF MONOCLONAL ANTIBODY EXPRESSION VECTOR

Publication Number WO/2020/135546
Publication Date 02.07.2020
International Application No. PCT/CN2019/128515
International Filing Date 26.12.2019
IPC
C12N 15/13 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
12Genes encoding animal proteins
13Immunoglobulins
C12N 15/85 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
C12N 15/11 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C07K 16/00 2006.01
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
16Immunoglobulins, e.g. monoclonal or polyclonal antibodies
CPC
C07K 16/00
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
16Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
C12N 15/11
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
11DNA or RNA fragments; Modified forms thereof
C12N 15/85
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09Recombinant DNA-technology
63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
79Vectors or expression systems specially adapted for eukaryotic hosts
85for animal cells
C12N 5/10
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
5Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor;
10Cells modified by introduction of foreign genetic material
Applicants
  • 先声生物医药科技有限公司 SIMCERE BIO-PHARMACEUTICAL TECHNOLOGY CO., LTD. [CN]/[CN]
  • 江苏先声药业有限公司 JIANGSU SIMCERE PHARMACEUTICAL CO., LTD. [CN]/[CN]
Inventors
  • 赵新燕 ZHAO, Xinyan
  • 卢士强 LU, Shiqiang
  • 赵东霞 ZHAO, Dongxia
  • 刘小青 LIU, Xiaoqing
  • 邓婧 DENG, Jing
  • 任晋生 REN, Jinsheng
Priority Data
201811618124.028.12.2018CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) METHOD FOR HIGH-THROUGHPUT CONSTRUCTION OF MONOCLONAL ANTIBODY EXPRESSION VECTOR
(FR) PROCÉDÉ DE CONSTRUCTION À HAUT RENDEMENT D'UN VECTEUR D'EXPRESSION D'ANTICORPS MONOCLONAL
(ZH) 高通量构建单克隆抗体表达载体的方法
Abstract
(EN)
Disclosed in the present invention is a method for the high-throughput construction of a monoclonal antibody expression vector and the application of said method in high-throughput antibody discovery, the method comprising the following steps: sorting antigen-specific single B cells into high-throughput reaction vessels, and implementing high-throughput reverse transcription; using the acquired reverse transcription reaction product as a template, respectively implementing PCR amplification of the heavy chain and light chain variable regions; and respectively recombining the heavy chain variable region clone fragments and light chain variable region clone fragments into an expression vector, the expression vector containing the suicide gene ccdB.
(FR)
La présente invention concerne un procédé de construction à haut rendement d'un vecteur d'expression d'anticorps monoclonal et l'application dudit procédé dans la découverte d'anticorps à haut rendement, le procédé consistant à trier des lymphocytes B uniques spécifiques d'un antigène dans des récipients de réaction à haut rendement, et mettre en oeuvre une transcription inverse à haut rendement; à utiliser le produit de réaction de transcription inverse obtenu comme modèle, mettant en oeuvre respectivement une amplification par PCR des régions variables de chaîne lourde et de chaîne légère; et à recombiner respectivement les fragments de clone de région variable de chaîne lourde et les fragments de clone de région variable de chaîne légère dans un vecteur d'expression, le vecteur d'expression contenant le gène suicide ccdB.
(ZH)
本发明公开了一种高通量构建单克隆抗体重组载体的方法以及该方法在高通量抗体发现中的应用,所述的方法包括以下步骤:分选抗原特异性单B细胞到高通量反应容器,并进行高通量逆转录;以获得的逆转录反应产物为模板,分别进行重链和轻链可变区的PCR扩增;将重链可变区克隆片段和轻链可变区克隆片段分别重组至表达载体中;所述的表达载体包含自杀基因ccdB。
Also published as
Latest bibliographic data on file with the International Bureau