Faculty Collaboration Database

Genetically detoxified tetanus toxin as a vaccine and conjugate carrier protein. Vaccine 2022 Aug 19;40(35):5103-5113

Date

07/26/2022

Pubmed ID

35871872

Pubmed Central ID

PMC10336728

DOI

10.1016/j.vaccine.2022.07.011

Scopus ID

2-s2.0-85134728035 (requires institutional sign-in at Scopus site)   6 Citations

Abstract

Tetanus toxoid (TTxd), developed over 100 years ago, is a clinically effective, legacy vaccine against tetanus. Due to the extreme potency of native tetanus toxin, manufacturing and regulatory efforts often focus on TTxd production, standardization, and safety, rather than product modernization. Recently, a genetically detoxified, full-length tetanus toxin protein (8MTT) was reported as a tetanus vaccine alternative to TTxd (Przedpelski et al. mBio, 2020). Here we describe the production of 8MTT in Gor/Met^TM E. coli, a strain engineered to have an oxidative cytoplasm, allowing for the expression of soluble, disulfide-bonded proteins. The strain was also designed to efficiently cleave N-terminal methionine, the obligatory start amino acid for E. coli expressed proteins. 8MTT was purified as a soluble protein from the cytoplasm in a two-column protocol to > 99 % purity, yielding 0.5 g of purified 8MTT/liter of fermentation broth with low endotoxin contamination, and antigenic purity of 3500 Lf/mg protein nitrogen. Mouse immunizations showed 8MTT to be an immunogenic vaccine and effective as a carrier protein for peptide and polysaccharide conjugates. These studies validate 8MTT as commercially viable and, unlike the heterogenous tetanus toxoid, a uniform carrier protein for conjugate vaccines. The development of a recombinant, genetically detoxified toxin produced in E. coli aligns the tetanus vaccine with modern manufacturing, regulatory, standardization, and safety requirements.

Author List

Chang MJ, Ollivault-Shiflett M, Schuman R, Ngoc Nguyen S, Kaltashov IA, Bobst C, Rajagopal SP, Przedpelski A, Barbieri JT, Lees A

Author

Joseph T. Barbieri PhD Professor in the Microbiology and Immunology department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Animals
Antibodies, Bacterial
Carrier Proteins
Escherichia coli
Mice
Tetanus
Tetanus Toxin
Tetanus Toxoid
Vaccines, Conjugate