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1. WO2020087988 - METHOD FOR SCREENING FUSION PROTEIN APTAMER AND KIT

Publication Number WO/2020/087988
Publication Date 07.05.2020
International Application No. PCT/CN2019/096330
International Filing Date 17.07.2019
IPC
C12N 15/10 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
10Processes for the isolation, preparation or purification of DNA or RNA
C12Q 1/6869 2018.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
6869Methods for sequencing
G01N 33/53 2006.01
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
CPC
C12N 15/1058
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
10Processes for the isolation, preparation or purification of DNA or RNA
1034Isolating an individual clone by screening libraries
1058Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
C12N 2320/13
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
2320Applications; Uses
10in screening processes
13in a process of directed evolution, e.g. SELEX, acquiring a new function
C12Q 1/6869
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6869Methods for sequencing
G01N 33/53
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
Applicants
  • 廖世奇 LIAO, Shiqi [CN]/[CN]
Inventors
  • 廖世奇 LIAO, Shiqi
  • 田彩平 TIAN, Caiping
  • 袁红霞 YUAN, Hongxia
  • 魏政丽 WEI, Zhengli
  • 栗怡 LI, Yi
Agents
  • 深圳市尔逊专利代理事务所(普通合伙) SHENZHEN ELSON PATENT AGENT FIRM(GENERAL PARTNERSHIP)
Priority Data
201811278341.X30.10.2018CN
Publication Language Chinese (ZH)
Filing Language Chinese (ZH)
Designated States
Title
(EN) METHOD FOR SCREENING FUSION PROTEIN APTAMER AND KIT
(FR) PROCÉDÉ DE CRIBLAGE D'APTAMÈRE DE PROTÉINE DE FUSION ET KIT
(ZH) 一种融合蛋白核酸适配体筛选方法及试剂盒
Abstract
(EN)
A method for screening a fusion protein aptamer and a kit in gene engineering. The method uses a two-way thermal cycling subtraction systematic evolution of ligands by exponential enrichment (SELEX), uses a fusion protein in gene engineering as a screening target and an epoxy (or carboxyl, NHS magnetic bead and streptomycin) agar magnetic bead as a vector, and conjugates a fusion tag protein antibody (or a biotinylated aptamer) onto the magnetic bead to form a capturing magnetic bead so as to capture the fusion tag protein expressed in gene engineering. The method also uses a random oligonucleotide library with a large capacity in combination with magnetic bead-tag protein antibody-tag protein-target protein to form a complex, thereby screening out the aptamer specifically binding to the target protein.
(FR)
Procédé de criblage d'un aptamère de protéine de fusion et kit en génie génétique. Le procédé utilise une évolution systématique par cyclage thermique à deux voies de ligands par enrichissement exponentiel (SELEX), une protéine de fusion en génie génétique en tant que cible de criblage et une bille magnétique époxy (ou un carboxyle, une bille magnétique NHS et de la streptomycine) pour agar en tant que vecteur, et associe un anticorps de protéine d'étiquette de fusion (ou un aptamère biotinylé) à une bille magnétique pour former une bille magnétique de capture de façon à capturer la protéine d'étiquette de fusion exprimée par génie génétique. Le procédé utilise en outre une vaste banque d'oligonucléotides aléatoires en combinaison avec une protéine étiquette-bille magnétique et un anticorps-protéine étiquette-protéine cible pour former un complexe, permettant ainsi de cribler l'aptamère se liant spécifiquement à la protéine cible.
(ZH)
一种基因工程中融合蛋白核酸适配体筛选方法及试剂盒。该方法利用双向热循环消减指数级富集的配体进化系统(SELEX),以基因工程中融合蛋白为筛选靶标,以环氧基(或羧基、NHS磁珠和链霉素)琼脂磁珠作为栽体,将融合标签蛋白抗体(或生物素化的核酸适配体)偶联到磁珠上,形成捕捉磁珠,捕获基因工程中表达的融合标签蛋白,利用大容量的随机寡核苷酸文库与磁珠-标签蛋白抗体-标签蛋白-靶蛋白结合形成复合物,从中筛选出与靶蛋白特异结合的核酸适配体。
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Latest bibliographic data on file with the International Bureau