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L2 multi-epitope fusion protein and application thereof

A technology of fusion protein and epitope, which is applied in the fields of application, hybrid peptide, medical preparations of non-active ingredients, etc.

Inactive Publication Date: 2014-09-24
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the modified IgG heavy chain protein is a very attractive vaccine carrier, and there is no report on its use in HPV vaccines

Method used

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  • L2 multi-epitope fusion protein and application thereof
  • L2 multi-epitope fusion protein and application thereof
  • L2 multi-epitope fusion protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Design and optimization of E3TR4 fusion gene

[0034] The upstream of the E3T gene contains the baculovirus gp67 signal peptide sequence, followed by 3 copies of the HPV16L2aa.17-36 epitope string, XhoI / KpnI and GGP linker sequences, and a general Th expression bit (SEQ ID NO: 3: AAFIAAATLKAAA). The structure of R4 gene is four copies of human IgG1 hinge region and CH2 domain (HCH2) tandem fusion with one copy of IgG1 Fc segment (including hinge region and CH2, CH3 domains). The E3T antigen gene and R4 are connected through a GGP linker to form a complete E3TR4 gene. The E3TR4 gene obtained by optimization of insect cell biased codons is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. E3TR4 gene structure diagram figure 1 shown.

[0035] The optimized E3TR4 gene sequence was synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd., and loaded into the pFastBac1 plasmid using EcoRI and HindIII restriction s...

Embodiment 2

[0036] Example 2: Construction of recombinant Bacmid of E3TR4 fusion gene and recombinant baculovirus

[0037] The pFastBac1-E3TR4 recombinant plasmid was used to transform Escherichia coli DH10Bac competent cells, and the recombinant Bacmid was obtained by screening, and then the recombinant Bacmid was transfected into insect cells Sf9, and the recombinant baculovirus was amplified in Sf9. The screening of recombinant Bacmid and the amplification method of recombinant baculovirus are well known, see for example patent CN101148661A.

Embodiment 3

[0038] Embodiment 3: the expression of E3TR4 fusion protein gene in Sf9 cell

[0039] Sf9 cells were inoculated with recombinant baculovirus to secrete and express the E3TR4 fusion antibody. After 88 hours, the culture supernatant was collected, centrifuged at 3000rpm for 15min, and the supernatant was collected for expression identification and purification. The method of infection expression is public, see for example patent CN101148661A.

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Abstract

The invention relates to an L2 multi-epitope fusion protein and application thereof. Specifically speaking, the invention relates to a fusion protein including concatermer of the L2 epitope of human papillomavirus (HPV) and universal auxiliary T lymphocytes and reconstructed mutant of an IgG subtype heavy-chain gene, coding nucleotide thereof, pFastBac1 recombinant plasmid including the coding nucleotide, recombinant Bacmid including the pFastBac1 recombinant plasmid, DH10Bac Escherichia coli including the recombinant Bacmid, an HPV prophylactic vaccine prepared from the above-mentioned components and application of the HPV prophylactic vaccine. The L2 multi-epitope fusion protein can be used for immunoprophylaxis of HPV infection and diseases related to HPV infection, and has important economic and social significance.

Description

technical field [0001] The invention relates to L2 multi-epitope fusion protein and application thereof. Specifically, the present invention relates to a fusion protein comprising a human papillomavirus L2 antigen protein sequence and a heavy chain protein sequence of an engineered antibody and its application. Background technique [0002] More than 100 types of human papillomavirus (HPV) have been isolated and identified, of which 1 / 3 are mucotropic viruses, including high-risk HPVs that induce malignant hyperplasia. , penile and oropharyngeal cancers are closely related to the occurrence of 15 types, and the positive rates detected in cervical cancer tissues from high to bottom are: HPV16, 18, 45, 31, 33, 52, 58, 35, 59, 56, 39, 51, 73, 68, 66; also includes 12 low-risk types that induce benign lesions such as condyloma acuminatum (HPV6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81 , 89), intermediate or suspected high-risk types (HPV34, 57, 83), and undetermined types (HPV34...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N15/866C12N1/21A61K39/12A61K47/48A61K48/00A61P31/20C12R1/19
Inventor 许雪梅陈雪刘洪洋张婷谢喜秀
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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