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SlyD-deficient Escherichia coli strains: A highway to contaminant-free protein extraction
Highlights: • Ε.coli ΔSlyD/SlyX strains engineered using λ-red mediated chromosomal deletion. • Development of several bacterial strains extends the spectrum of target proteins. • Preparation of SlyD/SlyX-free target protein after one purification step only. • ΔSlyD/SlyX bacterial strains allow to obtain a fully functional protein. Binding of native bacterial protein SlyD to metal affinity matrices remains a major problem in affinity purification of His-tagged recombinant proteins from Escherichia coli cells. In this study, four novel E. coli strains that lack the expression of SlyD/SlyX, were engineered using λ-red mediated chromosomal deletion. The resultant mutant E. coli strains allow us to obtain SlyD-free proteins immediately after metal affinity chromatography, and eliminate additional purification processes. As a model protein, bispecific antibodies composed of anti-F4/80 V{sub H}H module and anti-TNF V{sub H}H module (MYSTI-2) were used. Using this protein we have shown that the SlyD/SlyX-deficient E. coli strains allow us to obtain a fully functional protein.
Advancing phytomining: Harnessing plant potential for sustainable rare earth element extraction
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Thermodynamics of solvent-driven water extraction from hypersaline brines using dimethyl ether
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Microwave enhancement of extractions and reactions in Liquid-Liquid biphasic systems
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Intensification of solvent extraction in an additively manufactured microfluidic separator
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An Alternating Direction Minimization based denoising method for extracted ion chromatogram
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Depth sequence distribution of water extractable colloidal phosphorus and its phosphorus speciation
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Radiation Controllable Synthesis of Robust Covalent Organic Framework Conjugates for Efficient Dynamic Column Extraction of 99 TcO4 -
Wang et al. report here an ultra-robust imidazolium-decorated covalent organic framework (COF) conjugate fabricated by an ionizing radiation strategy for efficient capture of 99 TcO 4 -, where tunable ReO 4 - uptake up to 952 mg g -1 can be achieved by simply adjusting the γ-ray dose. The COFs feature high porosity, ultra-robust nanofiber structure, and fast adsorption kinetics, thus emphasizing their advantages in terms of column experiments.