Title:
METHOD FOR IMPROVING BIOCOUPLING EFFICIENCY BETWEEN PROTEIN AND NUCLEIC ACID BASED ON α-HELIX HANDLE
Document Type and Number:
WIPO Patent Application WO/2020/181756
Kind Code:
A1
Abstract:
Provided is a method for improving the biocoupling efficiency between a protein and a nucleic acid based on an α-helix handle. The method comprises designing a handle H-tag carrying an unnatural amino acid, then constructing a fusion recombinant expression plasmid containing the H-tag and a protein to be tested, expressing and purifying a fusion recombinant protein containing the unnatural amino acid in the H-tag, and finally subjecting the unnatural amino acid in the H-tag of the fusion protein and a coupling group on a nucleic acid substrate to efficient connection by means of click chemistry. The method uses an α-helix handle to provide controllable protein surface reaction conditions for an unnatural amino acid, and avoids a complex structure, charges and polar nano environment on the surface of the protein to be tested, thereby achieving the target of the efficient connection of a protein and a nucleic acid by means of click chemistry.
Inventors:
YU ZHONGBO (CN)
LI XU (CN)
MA XIAOFENG (CN)
QU LIHUA (CN)
HUANG WEI (CN)
LI XU (CN)
MA XIAOFENG (CN)
QU LIHUA (CN)
HUANG WEI (CN)
Application Number:
PCT/CN2019/107289
Publication Date:
September 17, 2020
Filing Date:
September 23, 2019
Export Citation:
Assignee:
UNIV NANKAI (CN)
International Classes:
C12N15/62; C07K19/00; C12N15/70
Foreign References:
| CN109750055A | 2019-05-14 | |||
| CN103764665A | 2014-04-30 | |||
| US20180333484A1 | 2018-11-22 |
Other References:
MORTEN, M. ET AL.: "Recent Advances in Covalent, Site-Specific Protein Immobilization", F1000RESEARCH, vol. 5, 12 September 2016 (2016-09-12), ISSN: 2046-1402
KIM, C.H. ET AL.: "Protein Conjugation with Genetically Encoded Unnatural Amino Acids", CURRENT OPINION IN CHEMICAL BIOLOGY, vol. 17, no. 3, 9 May 2013 (2013-05-09), XP055193458, ISSN: 1879-0402
ADDY, P.S. ET AL.: "A Chemoselective Rapid Azo-Coupling Reaction(CRACR) for ''Unclickable'' Bioconjugation", J. AM. CHEM. SOC., vol. 139, no. 34, 30 August 2017 (2017-08-30), XP055627975, ISSN: 1520-5126, DOI: 10.1021/jacs.7b05125
KIM, C.H. ET AL.: "Protein Conjugation with Genetically Encoded Unnatural Amino Acids", CURRENT OPINION IN CHEMICAL BIOLOGY, vol. 17, no. 3, 9 May 2013 (2013-05-09), XP055193458, ISSN: 1879-0402
ADDY, P.S. ET AL.: "A Chemoselective Rapid Azo-Coupling Reaction(CRACR) for ''Unclickable'' Bioconjugation", J. AM. CHEM. SOC., vol. 139, no. 34, 30 August 2017 (2017-08-30), XP055627975, ISSN: 1520-5126, DOI: 10.1021/jacs.7b05125
Attorney, Agent or Firm:
NUODE IP LAW FIRM (CN)
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