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Novel coronavirus N protein recombinant antigen and application thereof

A technology for recombinant antigen and protein recombination, which is applied in the direction of viruses, viral peptides, viruses/bacteriophages, etc., and can solve the problem of low thermal stability

Active Publication Date: 2020-10-30
SICHUAN MACCURA BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, in the current detection practice of the new coronavirus antibody, the full-length N protein still has some false positives as an antigen, and its thermal stability is low

Method used

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  • Novel coronavirus N protein recombinant antigen and application thereof
  • Novel coronavirus N protein recombinant antigen and application thereof
  • Novel coronavirus N protein recombinant antigen and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Sequence construction and expression purification of recombinant truncated N1 antigen

[0033] The truncated N1 antigen is a protein consisting of amino acids 25-214 of the N protein of the new coronavirus, and its amino acid sequence (SEQ ID NO.1) is as follows:

[0034] GSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSRNSSRNSTPGSSRGTSPARMAGNGG.

[0035] The gene expressing the truncated N1 antigen is a base-optimized nucleic acid named n1 gene, and its sequence (SEQ ID NO.2) is as follows:

[0036]

[0037] The n1 gene was constructed into the multiple cloning site of the pET28a vector to obtain a recombinant plasmid named pET28a-n1.

[0038] Transform pET28a-n1 into E. coli E. coli BL21 (Rossett) competent cells, and induce expression according to the method described in the Molecular Cloning Experiment Guide. The brief description is: 37°C...

experiment example 1

[0043] Experimental example 1 Comparison of binding properties of recombinant antigens

[0044] 1. Recombinant antigen

[0045] Truncated N1: Same as Example 1.

[0046] Truncated N2: select the 184-419th amino acid composition of the N protein of the new coronavirus, and the amino acid sequence (SEQ ID NO.3) is:

[0047] SRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMSSADSTQA。

[0048] Wild-type WN: wild-type N protein, the amino acid sequence (SEQ ID NO.4) is:

[0049] MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHIGTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLT...

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Abstract

The invention discloses a novel coronavirus N protein recombinant antigen and application thereof, and belongs to the field of protein immunoassay reagents. The recombinant antigen is a protein, and the amino acid sequence of the recombinant antigen is shown as SEQ ID NO.1 or 3. The recombinant antigen is used for preparing a reagent for detecting an N protein antibody; and compared with an N protein full-length antigen, the thermal stability of the reagent and the detection accuracy can be improved.

Description

technical field [0001] The invention belongs to the field of protein immunoassay reagents, and in particular relates to a novel coronavirus N protein recombinant antigen and its application. Background technique [0002] Coronavirus is a large virus family that exists widely in nature. It is named for its shape that looks like a crown under an electron microscope. It mainly causes respiratory diseases. The genome of the coronavirus is a single-stranded, positive-strand RNA with a total length of 26-32kb. Its genetic material is the largest among all RNA viruses, and it is divided into four genera: α, β, γ and δ. The infection of coronaviruses in nature is generally very widespread, and common mammals such as dogs, cats, mice, pigs, cattle, and poultry are susceptible. In recent years, various types of coronaviruses have been isolated from beluga whales, camels, and especially bats. There are currently six known human coronaviruses, namely HCoV-229E and HCoV-OC43 discovered...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/165C12N15/50G01N33/569G01N33/543
CPCC07K14/005C12N2770/20022G01N33/54326G01N33/56983G01N2333/165G01N2469/20
Inventor 李为民应斌武罗汶鑫王旻晋汪周峰
Owner SICHUAN MACCURA BIOTECH CO LTD
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