Clostridium perfringens epsilon toxin recombinant subunit vaccine and production method thereof

Abstract

The invention relates to a Clostridium perfringens epsilon toxin recombinant subunit vaccine and a production method thereof. The Clostridium perfringens epsilon toxin recombinant subunit vaccine prepared by the present invention is produced by codon-optimized recombinant Clostridium perfringens epsilon toxin protein containing 3 amino acid mutations, that is, the natural toxin protein is retained to the greatest extent Integrity and spatial conformation, thereby maintaining its immunogenicity, and avoiding the biological safety hazards caused by single amino acid mutations. The vaccine also has the advantages of simple preparation process, low immune dose, and much higher vaccine efficacy than existing vaccines. Compared with the current commercialized Clostridium perfringens natural toxin inactivated vaccine in China, the biological safety in the vaccine production process is greatly reduced. It is an ideal candidate vaccine for upgrading the current type D Clostridium perfringens toxin vaccine in my country; and when preparing a combined vaccine with other antigens, the combined vaccine can be prepared without increasing the dosage of the combined vaccine.

Description

technical field [0001] The invention relates to a Clostridium perfringens epsilon toxin recombinant subunit vaccine and a production method thereof. It belongs to the field of veterinary biological products. Background technique [0002] Clostridium perfringens, also known as Clostridium welchii, is an important zoonotic disease, the pathogen is traumatic gas gangrene and human food poisoning and sheep plague, lamb dysentery, cattle and sheep necrotic enteritis, cattle It is one of the main pathogens of sheep enterotoxemia, which has caused huge economic losses to animal husbandry. The main pathogenic factor of Clostridium perfringens is the exotoxin secreted by it. This bacterium can produce at least 18 kinds of exotoxins and play a pathogenic role through the toxins it produces. According to the types of four major lethal exotoxins α, β, ε and ι produced, the bacteria were divided into five toxin types A, B, C, D and E. Clostridium perfringens has the characteristics of...

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Problems solved by technology

[0006] Although there have been reports of attenuated mutants of Clostridium perfringens epsilon toxin, such as Yu Li et al. ("Clostridium perfringens epsilon toxin attenuated mutants and their applications", application No.: 201410707351.6) mutated the tyrosine at position 71 of the mature toxin of the wild-type Clostridium perfringens epsilon toxin into a non-aromatic amino acid, and obtained the epsilon toxin mutant attenuated for cells or animals. Since this application only Mutation of 1 amino acid is prone to reverse mutation and becomes a highly virulent toxin again, which will bring serious biosafety risks to the actual large-scale production of vaccines in the future

China Animal Disease Prevention and Control Center Song Xiaohui et al. ("Recombinant ε protein for inhibiting Clostridium perfringens infection and its preparation method and application", application number: 201610301989.9) designed 3 recombinant ε toxin proteins, but all of them lacked the wild type The first 50 amino acid residues of the epsilon toxin protein, this strategy of using protein fragments as immunogens is likely to result in incomplete antigenicity of the antigenic protein, thereby affecting its immunogenicity

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