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Basic Science and Pathogenesis.
Alzheimers Dement
December 2024
Emory University School of Medicine, Atlanta, GA, USA.
Background: The microtubule-associated Tau gene (MAPT) undergoes alternative splicing to produce isoforms with varying combinations of microtubule-binding region (MTBR) repeats (3R, 4R). The MTBR is the predominant region that forms paired helical filaments and neurofibrillary tangles fibrils in disease. Alzheimer's disease (AD) is a mixed Tauopathy containing both 3R and 4R isoforms.
View Article and Find Full Text PDFProtein purification with light via a genetically encoded azobenzene side chain.
Nat Commun
December 2024
Chair of Biological Chemistry, School of Life Sciences, Technical University of Munich, 85354, Freising, Germany.
Affinity chromatography is the method of choice for the rapid purification of proteins from cell extracts or culture supernatants. Here, we present the light-responsive Azo-tag, a short peptide comprising p-(phenylazo)-L-phenylalanine (Pap), whose side chain can be switched from its trans-ground state to the metastable cis-configuration by irradiation with mild UV light. Since only trans-Pap shows strong affinity to α-cyclodextrin (α-CD), a protein exhibiting the Azo-tag selectively binds to an α-CD chromatography matrix under daylight or in the dark but elutes quickly under physiological buffer flow when illuminating the column at 355 nm.
View Article and Find Full Text PDFEV-Elute: A universal platform for the enrichment of functional surface marker-defined extracellular vesicle subpopulations.
J Extracell Vesicles
December 2024
CDL Research, University Medical Center Utrecht, Utrecht, The Netherlands.
Intercellular communication via extracellular vesicles (EVs) has been identified as a vital component of a steadily expanding number of physiological and pathological processes. To accommodate these roles, EVs have highly heterogeneous molecular compositions. Given that surface molecules on EVs determine their interactions with their environment, EV functionality likely differs between subpopulations with varying surface compositions.
View Article and Find Full Text PDFImmobilizing calcium-dependent affinity ligand onto iron oxide nanoparticles for mild magnetic mAb separation.
Biotechnol Rep (Amst)
March 2025
Chair of Bioseparation Engineering, TUM School of Engineering and Design, Technical University of Munich, Boltzmannstraße 15, 85748 Garching, Germany.
Current downstream processing of monoclonal antibodies (mAbs) is limited in throughput and requires harsh pH conditions for mAb elution from Protein A affinity ligands. The use of an engineered calcium-dependent ligand (Z) in magnetic separation applications promises improvements due to mild elution conditions, fast processability, and process integration prospects. In this work, we synthesized and evaluated three magnetic nanoparticle types immobilized with the cysteine-tagged ligand Z-cys.
View Article and Find Full Text PDFSpyFixer enables efficient site-specific immobilization for protein-protein interaction analysis and antibody purification.
Int J Biol Macromol
December 2024
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong 510006, China. Electronic address:
Traditional methods of protein immobilization often result in activity loss due to random coupling. This study introduces SpyFixer, a variant of SpyCatcher that achieves over 99% efficient site-specific protein immobilization. We applied SpyFixer on two platforms: bio-layer interferometry (BLI) for protein-protein interaction analysis and epoxy agarose resin for antibody purification.
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