Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the ex...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-01-01
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| Series: | Toxins |
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| Online Access: | http://www.mdpi.com/2072-6651/11/1/35 |
| _version_ | 1811185870454128640 |
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| author | Bruna Xavier Rafaela Ferreira Perobelli Maurício Elesbão Walter Francielle Santos da Silva Sérgio Luiz Dalmora |
| author_facet | Bruna Xavier Rafaela Ferreira Perobelli Maurício Elesbão Walter Francielle Santos da Silva Sérgio Luiz Dalmora |
| author_sort | Bruna Xavier |
| collection | DOAJ |
| description | Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD50 mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use. |
| first_indexed | 2024-04-11T13:36:32Z |
| format | Article |
| id | doaj.art-852085f97269441ea0ce605ee1cc62e6 |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-11T13:36:32Z |
| publishDate | 2019-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-852085f97269441ea0ce605ee1cc62e62022-12-22T04:21:26ZengMDPI AGToxins2072-66512019-01-011113510.3390/toxins11010035toxins11010035Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and BioassaysBruna Xavier0Rafaela Ferreira Perobelli1Maurício Elesbão Walter2Francielle Santos da Silva3Sérgio Luiz Dalmora4Postgraduate Programme in Pharmaceutical Sciences; Federal University of Santa Maria, Santa Maria 97105-900, BrazilPostgraduate Programme in Pharmaceutical Sciences; Federal University of Santa Maria, Santa Maria 97105-900, BrazilPostgraduate Programme in Pharmaceutical Sciences; Federal University of Santa Maria, Santa Maria 97105-900, BrazilPostgraduate Programme in Pharmaceutical Sciences; Federal University of Santa Maria, Santa Maria 97105-900, BrazilIndustrial Pharmacy Department, Federal University of Santa Maria, Santa Maria 97105-900, BrazilBotulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD50 mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use.http://www.mdpi.com/2072-6651/11/1/35botulinum neurotoxin type Asize-exclusion chromatographyreversed-phase chromatographyT−47D cell cultureLD50 mouse bioassay |
| spellingShingle | Bruna Xavier Rafaela Ferreira Perobelli Maurício Elesbão Walter Francielle Santos da Silva Sérgio Luiz Dalmora Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays Toxins botulinum neurotoxin type A size-exclusion chromatography reversed-phase chromatography T−47D cell culture LD50 mouse bioassay |
| title | Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays |
| title_full | Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays |
| title_fullStr | Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays |
| title_full_unstemmed | Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays |
| title_short | Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays |
| title_sort | content potency assessment of botulinum neurotoxin type a by validated liquid chromatography methods and bioassays |
| topic | botulinum neurotoxin type A size-exclusion chromatography reversed-phase chromatography T−47D cell culture LD50 mouse bioassay |
| url | http://www.mdpi.com/2072-6651/11/1/35 |
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