Schistosoma japonica polypeptide with immunogenicity and application thereof

An immunogenic and schistosome technology, applied in the field of bioengineering, can solve the problems of no research reports and achieve good immune prevention effect

Inactive Publication Date: 2012-12-19
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there has been no research report on the application of receptor molecules that bind to the female groove protein of Schistosoma japonicum as a vaccine

Method used

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  • Schistosoma japonica polypeptide with immunogenicity and application thereof
  • Schistosoma japonica polypeptide with immunogenicity and application thereof
  • Schistosoma japonica polypeptide with immunogenicity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Preparation of anti-Schistosoma japonicum adult soluble antigen antibody

[0029] 1. Preparation of Soluble Antigens from Schistosoma japonicum Adult Worms

[0030] 1) New Zealand white rabbits were infected with 2000 cercariae of Schistosoma japonicum by abdominal patch method, and were killed after 42 days. The worms were collected by portal vein perfusion and frozen in liquid nitrogen for later use.

[0031] 2) Take out two tubes of Schistosoma japonicum worm bodies from liquid nitrogen, and lyse the worm bodies with PBS (1×PBS, pH 7.0, 0.5Mm PMSF, 0.1mM EDTA).

[0032] 3) Use a glass homogenizer to grind the worm bodies, put them in an ice bath for 15 minutes of ultrasonic lysis, and repeat freezing and thawing at -70°C three times.

[0033] 4) Ultrasonic cracking in an ice bath for 15 minutes.

[0034] 5) Centrifuge at 12000g for 1 hour, and collect the supernatant as adult soluble antigen (SWA).

[0035] 6) Detect the OD260 and OD280 values ​​of SWA w...

Embodiment 2

[0043] Example 2 Determination of titer of phage display cDNA library of adult Schistosoma japonicum (Sj44d)

[0044] The titer (titer) of phage is the number of live phage contained in one milliliter of culture medium. Phage counts can be performed because phages produce plaques visible to the naked eye on agar plates containing specific host bacteria. However, because the actual efficiency of the plaque counting method is difficult to approach 100% (generally low, because there are a few live phages that may not cause infection), in order to accurately express the concentration (titer or titer) of the virus suspension, it is generally not used The absolute number of virions is expressed in plaque-forming units (pfu for short). The specific steps of phage titer determination are as follows:

[0045] 1) Inoculate the host strain BLT5403 in 50ml of M9LB culture medium, shake at 37°C and 200rpm / min to make it OD 600nm The value reaches around 0.5;

[0046] 2) Take 10 μL of t...

Embodiment 3

[0050] Example 3 Screening of Schistosoma japonicum adult worm (Sj44d) phage display cDNA library

[0051] Based on the phage display cDNA library of Schistosoma japonicum adults (the phage display cDNA library of Schistosoma japonicum is preserved by our laboratory, and its library capacity is 4.98×10 6 pfu, the recombination rate is 93.8%), Schistosoma japonicum sclerotin was selected as a probe, and Schistosoma japonicum adult phage display cDNA library was screened in order to screen for receptor molecules related to S. The specific steps of method screening are as follows:

[0052] 1) Dilute the purified Schistosoma japonicum spore groove protein with TBS to a concentration of 1 μg / mL;

[0053] 3) Add 100 μL / well of Schistosoma japonicum dilute protein dilution to the ELISA plate, and coat overnight at 4°C;

[0054] 4) The next day, pour off the coating solution, pat dry, wash 5 times with TBST, shake on a shaker for 10 minutes each time;

[0055] 5) Add 150 μL of 3% B...

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Abstract

The invention discloses a schistosoma japonica polypeptide with immunogenicity, which can act with the gynecophoral canal protein of the schistosoma japonica mutually, and is provided with an amino acid sequence shown as SEQ ID NO: 1. The invention also discloses an application of the schistosoma japonica polypeptide with immunogenicity in preparing vaccines for preventing schistosomiasis or drugs for curing the schistosomiasis. The schistosoma japonica polypeptide with immunogenicity has a good immunoprophylaxis effect for the schistosomiasis, and is used as new vaccines for resisting the schistosomiasis.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to an immunogenic Schistosoma japonicum polypeptide and its application. Background technique [0002] Schistosomiasis is a parasitic disease that seriously endangers the health of humans and animals. Its susceptible animals include humans, cattle, sheep and other livestock and wild animals, involving more than 40 kinds of mammals. Livestock infection with schistosomiasis not only causes heavy losses to animal husbandry production, but also sick livestock is the main source of infection of human schistosomiasis in my country, and the situation of schistosomiasis prevention is still grim. Years of clinical practice have proved that chemotherapy can only temporarily reduce the incidence of schistosomiasis in areas where schistosomiasis has not been eradicated, but cannot completely block transmission. International experience shows that chemotherapy must be combined with a lon...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/435C12N15/12C12N7/01C12N1/21A61K39/00A61P33/12C07K16/18G01N33/68C12R1/19C12R1/92
CPCY02A50/30
Inventor 金亚美何金桃邹翔吴启进刘金明石耀军林矫矫
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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