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B cell epitope of Tarp protein of chlamydia trachomatis and application of B cell epitope

A Chlamydia trachomatis and B cell technology, applied in the fields of biomedical technology and immunology, can solve the problems of lack of prevention and treatment methods

Inactive Publication Date: 2015-04-29
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For Ct infectious diseases, there is still no effective prevention and treatment method so far, and seeking effective vaccines that can prevent and treat Ct infectious diseases is one of the important topics for workers in related disciplines in various countries.

Method used

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  • B cell epitope of Tarp protein of chlamydia trachomatis and application of B cell epitope
  • B cell epitope of Tarp protein of chlamydia trachomatis and application of B cell epitope
  • B cell epitope of Tarp protein of chlamydia trachomatis and application of B cell epitope

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1. Prediction and identification of Chlamydia trachomatis Tarp protein B cell epitope

[0065] 1. Design of Chlamydia trachomatis Tarp protein B cell epitope

[0066] Obtain the Chlamydia trachomatis (Ct) Tarp protein gene sequence and amino acid sequence from the network resource database (Genbank, Swiss-Prot); use the online network resource (EXPASY) and the biological software DNASTAR to analyze the antigenicity and surface accessibility of the Ct Tarp protein amino acid sequence , flexibility and transmembrane region and other parameters were analyzed ( figure 1 ), supplemented by the analysis results of the secondary structure of the CtTarp protein, analysis and comparison of various parameters to comprehensively judge the B cell dominant epitope of the CtTarp protein.

[0067] After the above epitope prediction, combined with repeated tests and selections by the inventors, the result was an amino acid sequence derived from the CtTarp protein, which is a p...

Embodiment 2

[0074] Example 2, Research on the Immunogenicity and Immunoprotection of Ct Tarp Protein B Cell Epitope Polypeptide

[0075] Female BALB / c mice aged 6-8 weeks were randomly divided into 3 groups, 6 mice in each group: the first group was the Ct Tarp protein B cell epitope immunization group, the second group was the KLH immunization control group, and the third group was the PBS control group. B cell epitope peptides were first coupled with KLH, and then the conjugates were fully emulsified with Freund's adjuvant (FCA) 1:1 (W / W), respectively, at 0, 2, and 4 weeks, 50 μg per mouse B cell epitope polypeptide, multi-point subcutaneous immunization of mice on the back. Tail vein blood and vaginal secretions were collected 2w, 4w, and 6w after immunization, and ELISA was used to detect polypeptide-specific IgG antibodies in serum and sIgA antibodies in secretions. The remaining mice in each group were challenged with E serotype Ct, and the activities of the mice and the severity...

Embodiment 3

[0085] Example 3, Antigenicity Research of CT Tarp Protein B Cell Epitope Polypeptide

[0086] 1. Immunoprecipitation and Western blot detection

[0087] Hela229 cells infected with Chlamydia for 48 hours were treated with cell lysate and then incubated with synthetic peptide immune serum for 4 hours. C overnight, then incubate with Protein G Agarose at 4°C for 3h, centrifuge, discard the supernatant, add the pellet to the loading buffer, perform SDS-PAGE protein electrophoresis, transfer to the membrane, and immunize mice with epitope peptides diluted 1:100 Serum was used as the primary antibody, HRP-labeled anti-mouse IgG (H+L) (purchased from eBioscience) was used as the secondary antibody, and the binding ability of B cell epitope immune serum to Tarp protein was detected by Western blot method. At the same time, Hela229 cells not infected with Chlamydia were used as negative control.

[0088] The results show( Figure 7 ), the B cell epitope protein immune serum can sp...

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Abstract

The invention belongs to the technical field of biology, and particularly discloses a B cell epitope which is obtained by screening on the basis of full-length Tarp protein of chlamydia trachomatis. The invention further discloses an encoding nucleotide and an amino acid sequence of the B cell epitope, and further discloses an application of the B cell epitope in prevention of chlamydia trachomatis infection diseases. The B cell epitope disclosed by the invention has the advantages of high immunogenicity and antigenicity, and good application prospect.

Description

technical field [0001] The invention belongs to the field of biomedicine technology and immunology, more specifically, the invention relates to a B cell epitope and its encoding nucleic acid from the Chlamydia trachomatis Tarp protein, and the epitope in the prevention of diseases related to Chlamydia trachomatis infection and / or application in medicine. Background technique [0002] Chlamydia trachomatis (Ct) is one of the main pathogens of sexually transmitted diseases (STDs). Mainly cause urethra in men and cervicitis in women after infection, it is reported that 50-60% of urethra in men is caused by Ct infection (Tiodorovi.J, Randelovi.G, Koci.B et al, Bacteriological finding in the urethra in men with and without non-gonococcal urethritis, 2007, 64(12): 833-6), and often mixed with other pathogens, infecting the female genitourinary tract through sexual transmission, leading to adverse pregnancy, such as miscarriage, stillbirth, etc., and is associated with cervical ca...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C12N15/31C07K16/12A61K39/118A61K39/40A61K48/00A61P31/04C12R1/01
Inventor 张丽芳朱珊丽薛向阳李文姝陈韶陈俊冯燕
Owner WENZHOU MEDICAL UNIV
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