期刊
SCIENTIFIC REPORTS
卷 12, 期 1, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41598-022-04931-7
关键词
-
资金
- Projekt DEAL
- Hessen State Ministry of Higher Education, Research and the Arts within the Hessen initiative for scientific and economic excellence (LOEWE-Program, LOEWE ZIB-Center for Insect Biotechnology and Bioresources)
The antibiotic resistance crisis has led to the study of alternative candidates, including antimicrobial peptides (AMPs). Continuous production processes have been found to be effective in meeting the demand for such molecules while achieving high yields. Process analytical technologies, like turbidimetry and dielectric spectroscopy, allow for online monitoring and control of cell growth, viability, and concentration. The perfusion process increased productivity by 180% compared to the batch process and achieved a high viable cell concentration.
The antibiotic resistance crisis has prompted research into alternative candidates such as antimicrobial peptides (AMPs). However, the demand for such molecules can only be met by continuous production processes, which achieve high product yields and offer compatibility with the Quality-by-Design initiative by implementing process analytical technologies such as turbidimetry and dielectric spectroscopy. We developed batch and perfusion processes at the 2-L scale for the production of BR033, a cecropin-like AMP from Lucilia sericata, in stably-transformed polyclonal Sf-9 cells. This is the first time that BR033 has been expressed as a recombinant peptide. Process analytical technology facilitated the online monitoring and control of cell growth, viability and concentration. The perfusion process increased productivity by similar to 180% compared to the batch process and achieved a viable cell concentration of 1.1 x 10(7) cells/mL. Acoustic separation enabled the consistent retention of 98.5-100% of the cells, viability was > 90.5%. The recombinant AMP was recovered from the culture broth by immobilized metal affinity chromatography and gel filtration and was able to inhibit the growth of Escherichia coli K12. These results demonstrate a successful, integrated approach for the development and intensification of a process from cloning to activity testing for the production of new biopharmaceutical candidates.
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