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Trivalent vaccine against human papilloma virus and its preparation and use

A human papillomavirus, virus-like technology

Active Publication Date: 2017-09-26
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In short, there is still a lack of low-cost, high-efficiency methods for preparing various HPV L1 proteins or VLPs in this field, so there is an urgent need to develop new methods for preparing HPV L1 proteins or VLPs with high efficiency, simplicity, and low cost, especially for the preparation of HPV L1 proteins or VLPs. A method to simultaneously contain L1 proteins or VLPs of HPV16, HPV18, and HPV58 to enable the development of vaccines against multiple specific subtypes

Method used

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  • Trivalent vaccine against human papilloma virus and its preparation and use
  • Trivalent vaccine against human papilloma virus and its preparation and use
  • Trivalent vaccine against human papilloma virus and its preparation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0184] Construction of high-efficiency expression system of HPV16L1 Pichia pastoris

[0185] 1. Optimization of HPV16L1 gene

[0186] Optimized sequence: According to the amino acid sequence of natural HPV16L1, the following factors are considered to optimize the design: the codon frequency used by Pichia pastoris, the G+C content in the gene and the secondary structure of mRNA.

[0187] The optimized nucleotide sequence of this embodiment is shown in SEQ ID NO:3, and the amino acid sequence of the HPV16L1 protein encoded by the sequence is shown in SEQ ID NO:2.

[0188] The optimized gene (SEQ ID NO: 3) was artificially synthesized, and BamHI and EcoRI restriction sites were introduced at its 5' end and 3' end respectively; the control sequence 1 used the wild-type HPV16L1 gene (SEQ ID NO: 1), Encoding the same HPV16L1 protein, after comparison, the homology between the optimized nucleic acid sequence and the control wild-type gene sequence is about 77.0%

[0189] 2. Kozak...

Embodiment 2

[0201] Construction of High Expression System of HPV18L1 Pichia pastoris

[0202] 1. Optimization of the target gene

[0203] Optimized sequence: According to the amino acid sequence of natural HPV18L1, the following factors are considered to optimize the design: the codon frequency used by Pichia pastoris, the G+C content in the gene, and the secondary structure of mRNA.

[0204] The nucleotide sequence of the target gene optimized in this example is shown in SEQ ID NO: 6, and the amino acid sequence of the HPV18L1 protein encoded by this sequence is identical to the natural sequence (shown in SEQ ID NO: 5). The optimized gene (SEQ ID NO: 6) was artificially synthesized, and BamHI and EcoRI restriction sites were introduced at its 5' end and 3' end respectively; the control sequence was the wild-type HPV18L1 gene (SEQ ID NO: 4), encoding For the same HPV18L1 protein, after comparison, the homology between the optimized sequence and the wild-type gene of the control is about...

Embodiment 3

[0211] Construction of High Expression System of HPV58L1 Pichia pastoris

[0212] Optimized sequence: According to the amino acid sequence of natural HPV58L1, the following factors are considered to optimize the design: the codon frequency used by Pichia pastoris, the G+C content in the gene, and the secondary structure of mRNA.

[0213] The optimized nucleotide sequence of this embodiment is shown in SEQ ID NO:9, and the amino acid sequence of the gene encoding HPV58L1 protein is shown in SEQ ID NO:8. The optimized gene (SEQ ID NO:9) was artificially synthesized, and BamHI and EcoRI restriction sites were introduced at its 5' end and 3' end, respectively.

[0214] Control sequence 1: wild-type HPV58L1 gene (SEQ ID NO: 7), encoding the same HPV58L1 protein; after comparison, the homology between the optimized sequence and the control wild-type gene is about 77.2%.

[0215] 2. Kozak sequence optimization

[0216] The optimized kozak sequence 1 screened in Example 1 was select...

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Abstract

The invention relates to a trivalent vaccine against human papillomavirus, its preparation method and application. Specifically, the present invention finds a kozak sequence that greatly improves the expression level of HPV L1 protein through a large number of screening and analysis of kozak sequences; Design and sequence optimization, eliminate the secondary structure of mRNA that affects translation, and construct a highly expressed HPV L1 gene expression cassette accordingly, which is especially suitable for expression in Pichia pastoris, and has high protein expression and stable Sexual characteristics.

Description

technical field [0001] The invention relates to the fields of genetic engineering and vaccinology, in particular, the invention relates to a trivalent vaccine against human papillomavirus and its preparation method and application. Background technique [0002] Cervical cancer is a common gynecological malignancy. Among the most common cancers in women, the incidence of cervical cancer ranks second. HPV (papillomavirus) virus infection is closely related to the occurrence of cervical cancer. At least 118 kinds of papillomaviruses have been isolated from humans so far. It has been confirmed that among anal and reproductive system cancers, cancers induced by HPV infection account for 3.7%. [0003] Among the HPV virus subtypes that have been isolated, 15 have been found to induce the occurrence of reproductive system cancer, so they are classified as high-risk HPV virus subtypes (such as HPV16, 18, 31, 33, 52 and 58), Two subtypes of HPV16 and HPV18 account for 70% of all ce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/37C12N15/63C12N1/21C12N1/19C12N5/10C12P21/02C12N7/04A61K39/12A61P31/20C12R1/84
Inventor 潘卫庆徐新东张远斌
Owner TONGJI UNIV
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