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Single-Walled Carbon Nanotube Probes for Protease Characterization Directly in Cell-Free Expression Reactions.
bioRxiv
January 2025
Chemical and Biological Engineering - Iowa State University, 618 Bissell Rd, Ames, IA 50011.
Proteins can be rapidly prototyped with cell-free expression (CFE) but in most cases there is a lack of probes or assays to measure their function directly in the cell lysate, thereby limiting the throughput of these screens. Increased throughput is needed to build standardized, sequence to function data sets to feed machine learning guided protein optimization. Herein, we describe the use of fluorescent single-walled carbon nanotubes (SWCNT) as effective probes for measuring protease activity directly in cell-free lysate.
View Article and Find Full Text PDFBackground: Deficiency in the lysosomal enzyme, glucocerebrosidase (GCase), caused by mutations in the GBA1 gene, is the most common genetic risk factor for Parkinson's disease (PD). However, the consequence of reduced enzyme activity within neural cell sub-types remains ambiguous. Thus, the purpose of this study was to define the effect of GCase deficiency specifically in human astrocytes and test their non-cell autonomous influence upon dopaminergic neurons in a midbrain organoid model of PD.
View Article and Find Full Text PDFPreparation and application of a multiepitope fusion protein based on bioinformatics and Tandem Mass Tag-based proteomics technology.
Front Immunol
January 2025
Jiangsu Engineering Research Center of Biological Data Mining and Healthcare Transformation, Xuzhou Medical University, Xuzhou, China.
Introduction: Brucellosis is a widespread zoonotic disease that poses a considerable challenge to global public health. Existing diagnostic methods for this condition, such as serological assays and bacterial culture, encounter difficulties due to their limited specificity and high operational complexity. Therefore, there is an urgent need for the development of enhanced diagnostic approaches for brucellosis.
View Article and Find Full Text PDFPolysorbates degrading enzymes in biotherapeutics - a current status and future perspectives.
Front Bioeng Biotechnol
January 2025
Pharmaceutical Development Biologicals, TIP, Boehringer Ingelheim Pharma GmbH & Co., KG, Innovation Unit, Biberach an der Riss, Germany.
Polysorbates, in particular polysorbate (PS) 20 and 80, are the most commonly used surfactants for stabilising biotherapeutics produced by biotechnological processes. PSs are derived from ethoxylated sorbitan (a derivative of sorbitol) esterified with fatty acids of varying chain length and degree of saturation. In the past, these surfactants have been reported to have specific liabilities.
View Article and Find Full Text PDFCharacterization of the GH72 family of Laminarin/Lichenin transferases and their roles in cell wall biogenesis.
Cell Surf
June 2025
Department of Biological Sciences, SUNY University at Buffalo, Buffalo, NY 14260, United States.
In vegetative hyphae, chitin, β-1,3-glucan (laminarin), and a mixed β-1,3-/β-1,4-glucan (lichenin) are the major cell wall polysaccharides. GH72 enzymes have been shown to function as β-1,3-glucanases and glucanosyltransferases and can function in crosslinking β-1,3-glucans together. To characterize the enzymatic activities of the enzymes, we expressed GEL-1 with a HIS6 tag in A chimeric maltose binding protein:GEL-2 was produced in .
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