Optimization of tetanus toxoid ammonium sulfate precipitation process using response surface methodology

Marchetti-Deschmann, Martina and Ranić, Jelena and Kurtović, Tihana and Prebeg, Pero and Brgles, Marija and Allmaier, Günter and Halassy, Beata (2016) Optimization of tetanus toxoid ammonium sulfate precipitation process using response surface methodology. = Optimization of tetanus toxoid ammonium sulfate precipitation process using response surface methodology. Preparative Biochemistry and Biotechnology, 46 (7). pp. 695-703. ISSN 1082-6068. Vrsta rada: ["eprint_fieldopt_article_type_article" not defined]. Kvartili JCR: Q3 (2016). Točan broj autora: 7.

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Abstract

Tetanus toxoid (TTd) is a highly immunogenic, detoxified form of tetanus toxin, a causative agent of tetanus disease, produced by Clostridium tetani. Since tetanus disease cannot be eradicated but is easily prevented by vaccination, the need for the tetanus vaccine is permanent. The aim of this work was to investigate the possibility of optimizing TTd purification i.e. ammonium sulfate precipitation process. The influence of the percentage of ammonium sulfate, starting amount of TTd, buffer type, pH, temperature and starting purity of TTd on the purification process were investigated using optimal design for response-surface models. Responses measured for evaluation of the ammonium sulfate precipitation process were TTd amount (Lf/mL) and total protein content. These two parameters were used to calculate purity (Lf/mgPN) and the yield of the process. Results indicate that citrate buffer, lower temperature and lower starting amount of TTd result in higher purities of precipitates. Gel electrophoresis combined with MALDI mass spectrometric analysis of precipitates revealed that there are no inter-protein cross-links and that all contaminating proteins have pIs similar to TTd so this is most probably the reason for the limited success of purification by precipitation.

Item Type: Article (["eprint_fieldopt_article_type_article" not defined])
Keywords (Croatian): ammonium sulfate precipitation, Design of Experiments, mass spectrometry, radial immunodiffusion, tetanus toxoid, tetanus vaccine
Subjects: TECHNICAL SCIENCE
Divisions: 600 Department of Naval Engineering and Marine Technology > 620 Chair of Marine Structures Design
Indexed in Web of Science: Yes
Indexed in Current Contents: Yes
Quartiles: Q3 (2016)
Date Deposited: 23 May 2016 11:03
Last Modified: 19 Mar 2018 12:09
URI: http://repozitorij.fsb.hr/id/eprint/5917

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