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Proteolysis is one of the reasons for poor production of recombinant proteins in Escherichia coli. Important properties of E. coli proteases, which are relevant for the production of recombinant proteins, are reviewed. Furthermore, various strategies to control the proteolysis of the recombinant proteins are presented. These strategies for control of proteolysis can be applied on various stages of the process: design of more stable protein, a modification of the host cell in respect to proteolytic activity, optimisation of cultivation and downstream processing. However, before implementing these measures the proteolysis rate should be measured in order to calculate a potential benefit of reduced proteolysis rate.
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http://dx.doi.org/10.1007/b95567 | DOI Listing |
Appl Biochem Biotechnol
September 2025
School of Biological Sciences, University of the Punjab, Quaid-E-Azam Campus, P.O. 54590, Lahore, Pakistan.
Recombinant DNA technology is widely used to produce industrially and pharmaceutically important proteins. In silico analysis, performed before executing wet lab experiments has been greatly helpful in this connection. A shift in protein analysis has been observed over the past decade, driven by advancements in bioinformatics databases, tools, software, and web servers.
View Article and Find Full Text PDFElife
September 2025
Chinese Academy of Medical Science Oxford Institute, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom.
Influenza virus neuraminidase (NA) is a crucial target for protective antibodies, yet the development of recombinant NA protein as a vaccine has been held back by instability and variable expression. We have taken a pragmatic approach to improving expression and stability of NA by grafting antigenic surface loops from low-expressing NA proteins onto the scaffold of high-expressing counterparts. The resulting hybrid proteins retained the antigenic properties of the loop donor while benefiting from the high-yield expression, stability, and tetrameric structure of the loop recipient.
View Article and Find Full Text PDFACS Synth Biol
September 2025
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, P. R. China.
Human Bone Morphogenetic Protein-2 (hBMP-2) serves as a critical regulator in bone and cartilage formation; however, its industrial application is hindered by its inherent tendency to form inclusion bodies in prokaryotic expression systems. To address this issue, we established a recombinant hBMP-2 (rhBMP-2) expression system using the pCold II plasmid and the SHuffle T7 strain. We explored several strategies to enhance the solubility of rhBMP-2, including coexpression with molecular chaperones, vesicle-mediated secretory expression, fusion expression with synthetic intrinsically disordered proteins (SynIDPs), and fusion expression with small-molecule peptide tags.
View Article and Find Full Text PDFAllergol Immunopathol (Madr)
September 2025
Faculty of Medicine, University of Prishtina, University Clinical Center of Kosovo, Prishtina, Republic of Kosovo.
Objective: The aim of this study was to assess the association between allergic reactions after COVID-19 vaccination and the history of high-risk allergy, individual predisposing factors such as age and gender, and COVID-19 vaccine type.
Materials And Methods: This retrospective cohort study included 234 adult patients (18 years old and above) who underwent a COVID-19 vaccine allergy test up until February 2023 in a Clinic of Allergy and Clinical Immunology in the University Clinical Center of Kosovo. All patients suspected of allergy underwent skin testing: SPT (skin prick test) and IDT (intradermal test) using either an mRNA (ribonucleic messenger acid) vaccine (BNT162b2, Pfizer-BioNTech) and/or an adenoviral vector vaccine (AZD1222, AstraZeneca).
J Proteome Res
September 2025
Center for Proteomics and Metabolomics, Leiden University Medical Center, Postbus 9600, 2300 RC Leiden, The Netherlands.
Plasma proteomics has regained attention in recent years through advancements in mass spectrometry instrumentation and sample preparation as well as new high-throughput affinity-based technologies. Here, we evaluate the analytical performance of the new Olink Reveal platform, a proximity extension assay (PEA)-based technology quantifying 1034 proteins and covering many biological pathways, in particular immune system processes. Using spiked-in recombinant Interleukin-10 (IL-10) and vascular endothelial growth factor D (VEGF-D) in the NIST SRM 1950 plasma standard, we assessed the linearity, sensitivity, precision, and accuracy of the Olink Reveal assay.
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