ArticleEngineering in life sciences2017
Design of experiments-based high-throughput strategy for development and optimization of efficient cell disruption protocols.
Article in Engineering in life sciences, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 32 citations in OpenAlex.
- MATRIX: rapid quantification of total and active microbial cells with single-cell phenotypes for environmental microbiomes.mSystems · 2026Article
- Lysis and IB washing procedures significantly impact the recovery of rHumanized ranibizumab from inclusion bodies expressed in Escherichia coli.Bioprocess and biosystems engineering · 2026Article
- Characterizing and Tailoring the Substrate Profile of a γ-Glutamyltransferase Variant.ACS synthetic biology · 2024Article
- Characterization of putative mannoprotein inSynthetic and systems biotechnology · 2023Article
- Model-assisted DoE software: optimization of growth and biocatalysis in Saccharomyces cerevisiae bioprocesses.Bioprocess and biosystems engineering · 2021Article
- Monte Carlo Simulations for the Analysis of Non-linear Parameter Confidence Intervals in Optimal Experimental Design.Frontiers in bioengineering and biotechnology · 2019Article
- Accelerated Bioprocess Development of Endopolygalacturonase-Production withBioengineering (Basel, Switzerland) · 2018Article
- Data-driven multi-objective optimization via grid compatible simplex technique and desirability approach for challenging high throughput chromatography applications.Biotechnology progress · 2018Article
- Automated Cell Treatment for Competence and Transformation ofMicroorganisms · 2018Article
- Online bioprocess data generation, analysis, and optimization for parallel fed-batch fermentations in milliliter scale.Engineering in life sciences · 2017Article
- Editorial: Bioprocess Development in the era of digitalization.Engineering in life sciences · 2017Article
- Artificial Intelligence vs. Statistical Modeling and Optimization of Continuous Bead Milling Process for Bacterial Cell Lysis.Frontiers in microbiology · 2016Article
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Efficient and reproducible cell lysis is a crucial step during downstream processing of intracellular products. The composition of an optimal lysis buffer should be chosen depending on the organism, its growth status, the applied detection methods, and even the target molecule. Especially for high-throughput applications, where sample volumes are limited, the adaptation of a lysis buffer to the specific campaign is an urgent need. Here, we present a general design of experiments-based strategy suitable for eight constituents and demonstrate the strength of this approach by the development of an efficient lysis buffer for Gram-negative bacteria, which is applicable in a high-throughput format in a short time. The concentrations of four lysis-inducing chemical agents EDTA, lysozyme, Triton X-100, and polymyxin B were optimized for maximal soluble protein concentration and ß-galactosidase activity in a 96-well format on a Microlab Star liquid handling platform under design of experiments methodology. The resulting lysis buffer showed the same performance as a commercially available lysis buffer. The developed protocol resulted in an optimized buffer within only three runs. The established procedure can be easily applied to adapt the lysis buffer to other strains and target molecules.
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