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Amino acid sequence of recombined human papilloma virus L1 capsid protein and uses thereof

A technology for human papilloma virus and capsid protein, which is applied in the preparation of vaccines, the application of diagnosing antigens or antibodies, the HPV L1 protein pentamer, and the fields of pharmaceutical compositions, which can solve the difficulties of biological activity expression, difficulty in activity expression, and cost High-level problems, to achieve major economic and social effects, convenient quality control, and low cost

Inactive Publication Date: 2008-08-20
马润林 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, prokaryotic expression technology also has defects: the active expression of biological macromolecules with large molecular weight is relatively difficult, especially the biological activity expression of many genes in higher animals and humans is more difficult
In order to produce some polymorphic drugs or biological agents with important biological activities, people have to adopt the costly and complicated eukaryotic fermentation method

Method used

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  • Amino acid sequence of recombined human papilloma virus L1 capsid protein and uses thereof
  • Amino acid sequence of recombined human papilloma virus L1 capsid protein and uses thereof
  • Amino acid sequence of recombined human papilloma virus L1 capsid protein and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Taking HPV16 as an example to illustrate the recombination of the amino acid sequence of HPV L1 capsid protein

[0046] 1. Detection and isolation of human papillomavirus type 16 (HPV16) DNA: The waste of clinical cell samples that may contain wild-type HPV16 virus (the remaining part of cervical exfoliated cells used for routine cytomorphological examination) was purchased from Beijing An Gynecology Clinic of Zhen Hospital. Suspended in 1.0 ml of water (about 1×10 5 1) exfoliated cells were treated with 1000 units of proteinase K (purchased from Huamei Biological Products Co., Ltd.) at 55 degrees Celsius for 300 minutes, and centrifuged at 10000 g for 30 minutes at a high speed. The resulting supernatant can be used to detect whether there is HPV virus. The infected HPV16 positive cell supernatant can be directly used for PCR amplification of the target gene fragment of the HPV16 virus DNA sequence.

[0047] 2. Molecular cloning of HPV16 L1 protein DNA: HPV...

Embodiment 2

[0054] Example 2 Recombination Transformation of the Amino Acid Sequence of the L1 Capsid Protein of the Other HPV Types

[0055] Recombination transformation of the amino acid sequence of L1 capsid protein of other HPV types (including HPV6, HPV11, HPV18, HPV26, HPV31, HPV33, HPV35, HPV39, HPV42, HPV45, HPV 51, HPV 52, HPV 53, HPV 56, HPV58, HPV 59. The operation steps and detection methods of HPV 66, HPV 73, and HPV 82) are the same as those in Example 1, and the differences are shown in Table 1.

[0056] Table 1

[0057] HPV type

[0058] HPV26

[0059] After the amino acid sequence of the L1 capsid protein of the HPV type was recombined, the results obtained were the same as those of the HPV16 L1 capsid protein obtained in Example 1, that is, a water-soluble L1 pentamer was obtained.

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Abstract

The invention relates to an amino acid sequence of a recombinant human papillomavirus L1 capsid protein, a nucleotide sequence which encodes the amino acid sequence and a recombinant vector and an expression host which contain the nucleotide sequence. The invention further relates to an application of a HPV L1 protein which is composed of the amino acid sequence in the preparation of vaccines, drug combination and diagnostic antigens or antibodies. The invention allows the recombinant HPV L1 capsid protein which is expressed in a prokaryotic system to be dissolvable in water by the modification of the HPV L1 wild-type sequence, and an L1 pentamer which has the same immunogenicity and antigenicity with the VLP of HPV L1 is obtained by expression. The invention allows the industrial production of the HPV L1 capsid protein by utilizing the prokaryotic expression system to become a reality, compared with the currently used eukaryotic expression system, the invention has the advantages of more stable quality of the products, higher yield, low cost and convenient quality control, which has great economic benefits and social effects.

Description

technical field [0001] The present invention relates to the field of prevention and treatment of human papillomavirus infection. Specifically, the present invention relates to the amino acid sequence of recombinant human papillomavirus L1 capsid protein, the nucleotide sequence encoding the amino acid sequence, and the vector and host cell comprising these nucleotide sequences. The application of the formed HPV L1 protein pentamer in the preparation of vaccines, pharmaceutical compositions, diagnostic antigens or antibodies. Background technique [0002] Papillomaviruses are a large group of DNA viruses that infect humans and other animals. Among them, Human papilloma virus (HPV) is associated with various human diseases, including benign warts and cancers. [0003] Papillomaviruses contain two structural proteins (or capsid proteins) for protecting viral chromosomes, L1 and L2 respectively. 360 L1 protein molecules or 72 L1 pentamers form the shell of the virus particle; ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/025C12N15/37C12N15/63C12N1/21A61K39/12A61P31/12G01N33/53G01N33/68G01N33/569
CPCC12N2710/20034A61K39/12A61K2039/645C12N2710/20022C07K14/005G01N2333/025C12N7/00G01N33/56983A61P31/12A61P31/20
Inventor 马润林陈小江
Owner 马润林
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