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

Autor: Alves GG; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil., Assis RA; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil., do Amarante VS; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil., de Oliveira Júnior CA; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil., Silva ROS; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil. Electronic address: rodrigo.otaviosilva@gmail.com., Dias Heneine LG; Fundação Ezequiel Dias, Rua Conde Pereira Carneiro, 80 - Gameleira, Belo Horizonte, 30510-010, MG, Brazil., Lobato FCF; Veterinary School, Universidade Federal de Minas Gerais (UFMG), Avenida Antônio Carlos, 6627, Belo Horizonte, MG, CEP 31.270-901, Brazil. Electronic address: lobato.francisco@yahoo.com.br.
Jazyk: angličtina
Zdroj: Anaerobe [Anaerobe] 2024 Feb; Vol. 85, pp. 102817. Date of Electronic Publication: 2023 Dec 30.
DOI: 10.1016/j.anaerobe.2023.102817
Abstrakt: 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.
Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest.
(Copyright © 2023 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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