The potential for developing improved procedures for phosphate measurement through combinations of gel electrophoresis and quadrupole-based ICP mass spectrometry utilising (47)PO(+) is investigated. Laser ablation of gels offers a rapid and direct quantitation route, but is subject to high blanks due to P impurities in gels and associated reagents; nevertheless optimisation of laser sampling afforded improved method sensitivity (limit of detection 0.09 microg g(-1)). Implementation of whole gel elution (WGE) with FI-ICP-MS (conventional solution nebulisation) following gel electrophoresis permitted quantitation at the sub microg l(-1) level, and microcolumn processing (activated alumina) was effective at rejecting phosphate contamination. The potential for S-induced molecular ion interference at mass 47 was demonstrated.
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http://dx.doi.org/10.1007/s00216-005-3361-7 | DOI Listing |
J Proteome Res
January 2025
Impact Proteomics, LLC., Pittsburgh, Pennsylvania 15206, United States.
Immunoprecipitation is among the most widely utilized methods in biomedical research, with applications that include the identification of antibody targets and associated proteins. The path to identifying these targets is not straightforward, however, and often requires the use of chemical cross-linking and/or gel electrophoresis to separate targets from an overabundance of immunoglobulin protein. Such experiments are labor intensive and often yield long lists of candidate antibody targets.
View Article and Find Full Text PDFBiochimie
January 2025
Department of Biochemistry, School of Biology, M.V. Lomonosov Moscow State University; Department of Biochemistry and Regenerative Biomedicine Faculty of Basic Medicine, M.V. Lomonosov Moscow State University. Electronic address:
BAG3 is a universal adapter protein involved in various cellular processes, including the regulation of apoptosis, chaperone-assisted selective autophagy, and heat shock protein function. The interaction between small heat shock proteins (sHsps) and their α-crystallin domains (Acds) with full-length BAG3 protein and its IPV domain was analyzed using size-exclusion chromatography, native gel electrophoresis, and chemical cross-linking. HspB7 and the 3D mutant of HspB1 (which mimics phosphorylation) showed no interaction, HspB6 weakly interacted, and HspB8 strongly interacted with full-length BAG3.
View Article and Find Full Text PDFACS Nano
January 2025
State Key Laboratory of Organic Electronics and Information Displays & Institute of Advanced Materials (IAM), National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing University of Posts and Telecommunications, 9 Wenyuan Road, Nanjing 210023, China.
Higher-order DNA nanomaterials have emerged as programmable tools for probing biological processes, constructing metamaterials, and manipulating mechanically active nanodevices with the multifunctionality and high-performance attributes. However, their utility is limited by intricate mixtures formed during hierarchical multistage assembly, as standard techniques like gel electrophoresis lack the resolution and applicability needed for precise characterization and enrichment. Thus, it is urgent to develop a sorter that provides high separation resolution, broad scope, and bioactive functionality.
View Article and Find Full Text PDFCurr Pharm Des
January 2025
School of Chemical Engineering, College of Engineering, University of Tehran, Tehran, Iran.
Hemophilia A (HA) is an inherited condition that is characterized by a lack of coagulation factor VIII (FVIII), which is needed for blood clotting. To produce recombinant factor VIII (rFVIII) for treatment, innovative methods are required. This study presents a thorough examination of the genetic engineering and biotechnological methods that are essential for the production of this complex process.
View Article and Find Full Text PDFJ Pharm Biomed Anal
January 2025
State Key Laboratory of Neurology and Oncology Drug Development, Nanjing, China; Simcere Zaiming Pharmaceutical Co, Ltd., Nanjing, China. Electronic address:
Capillary electrophoresis-sodium dodecyl sulfate (CE-SDS) is widely used in the biopharmaceutical industry for monitoring purity and analyzing impurities. The accuracy of the method may be compromised by artificial species resulting from sample preparation or electrophoresis separation due to suboptimal conditions. During non-reduced CE-SDS analysis of a multispecific antibody (msAb), named as multispecific antibody C (msAb-C), a cluster of unexpected peaks was observed after the main peak.
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