Production and purification of Clostridium perfringens type C beta-toxin and IgG and IgY antitoxins

IF 2.5 3区生物学 Q3 MICROBIOLOGY Anaerobe Pub Date : 2023-12-30 DOI:10.1016/j.anaerobe.2023.102817

Guilherme Guerra Alves , Ronnie Antunes Assis , Victor Santos do Amarante , Carlos Augusto de Oliveira Júnior , Rodrigo Otávio Silveira Silva , Luiz Guilherme Dias Heneine , Francisco Carlos Faria Lobato

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Abstract

Objectives

This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY).

Methods

Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY.

Results

All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %).

Conclusions

C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.

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生产和纯化 C 型产气荚膜梭菌 beta 型毒素以及 IgG 和 IgY 抗毒素

目的本研究旨在生产和纯化 C 型产气荚膜梭菌 beta-毒素、绵羊抗 beta-毒素免疫球蛋白 G (IgG) 和鸡免疫球蛋白 Y (IgY)。方法采用两种方法纯化 beta-毒素：以锌为螯合剂的单步金属亲和层析法 (MAC) 和离子交换层析法 (IEX)。然后将纯化和灭活的 beta 毒素注射给羊和鸡，以产生 IgG 和 IgY。相比之下，MAC 在单步方案中每次运行可纯化超过 21 毫克的毒素。然后将纯化和灭活的 C 型毒素给绵羊和鸡注射，纯化出的 IgG 和 IgY 产率高，抗体滴度为 50 IU/mL，亲和力高（73.2%）。该方案可用于生产诊断和研究 C 型产气荚膜杆菌引起的坏死性肠炎的成分，也可用于评估现有疫苗和开发预防该疾病的新方法。

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来源期刊

Anaerobe 生物-微生物学

CiteScore

5.20

自引率

8.70%

发文量

137

审稿时长

76 days

期刊介绍： Anaerobe is essential reading for those who wish to remain at the forefront of discoveries relating to life processes of strictly anaerobes. The journal is multi-disciplinary, and provides a unique forum for those investigating anaerobic organisms that cause infections in humans and animals, as well as anaerobes that play roles in microbiomes or environmental processes. Anaerobe publishes reviews, mini reviews, original research articles, notes and case reports. Relevant topics fall into the broad categories of anaerobes in human and animal diseases, anaerobes in the microbiome, anaerobes in the environment, diagnosis of anaerobes in clinical microbiology laboratories, molecular biology, genetics, pathogenesis, toxins and antibiotic susceptibility of anaerobic bacteria.