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Proteomics techniques in protein biomarker discovery

Quantitative Biology 2024年第1期12卷 53-69页

作者： Mahsa Babaei Soheila Kashanian Huang-Teck Lee frances harding Department of Biology Faculty of ScienceRazi UniversityKermanshahIran Faculty of Chemistry Sensor and Biosensor Research Center(SBRC)Razi UniversityKermanshahIran Nanobiotechnology Department Faculty of Innovative Science and TechnologyRazi UniversityKermanshahIran Centre for Marine Bioproducts Development Flinders UniversityAdelaideSouth AustraliaAustralia Monash Institute of Pharmaceutical Sciences Monash UniversityMelbourneNew South WalesAustralia

Protein biomarkers represent specific biological activities and processes, so they have had a critical role in cancer diagnosis and medical care for more than 50 years. With the recent improvement in proteomics technologies, thousands of protein biomarker candidates have been developed for diverse disease states. Studies have used different types of samples for proteomics diagnosis. Samples were pretreated with appropriate techniques to increase the selectivity and sensitivity of the downstream analysis and purified to remove the contaminants. The purified samples were analyzed by several principal proteomics techniques to identify the specific protein. In this study, recent improvements in protein biomarker discovery, verification, and validation are investigated. Furthermore, the advantages, and disadvantages of conventional techniques, are discussed. Studies have used mass spectroscopy (MS) as a critical technique in the identification and quantification of candidate biomarkers. Nevertheless, after protein biomarker discovery, verification and validation have been required to reduce the false-positive rate where there have been higher number of samples. Multiple reaction monitoring (MRM), parallel reaction monitoring (PRM), and selected reaction monitoring (SRM), in combination with stable isotope-labeled internal standards, have been examined as options for biomarker verification, and enzyme-linked immunosorbent assay (ELISA) for validation.

关键词： biomarker discovery cancer biomarker gel-based methods gel-free methods mass spectroscopy proteomics

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Oligonucleotide delivery by chitosan-functionalized porous silicon nanoparticles

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Nano Research 2015年第6期8卷 2033-2046页

作者： Morteza Hasanzadeh Kafshgari Bahman Delalat Wing Yin Tong frances J. harding Martti Kaasalainen Jarno Salonen Nicolas H. Voelcker ARC Centre of Excellence in Convergent Bio-Nano Science and Technology Mawson Institute University of South Australia GPO Box 247 Adelaide SA 500 1 Australia Department of Physics and Astronomy University of Turku FI-20014 Turku Finland

Porous silicon nanoparficles （pSiNPs） are a promising nanocarrier system for drug delivery owing to their biocompatibility, biodegradability, and non-inflammatory nature. Here, we investigate the fabrication and characterization of thermally hydrocarbonized pSiNPs （THCpSiNPs） and chitosan-coated THCpSiNPs for therapeutic oligonucleotide delivery. Chitosan coating after oligonucleotide loading significantly improves sustained oligonucleotide release and suppresses burst release effects. Moreover, cellular uptake, endocytosis, and cytotoxicity of oligonucleotide-loaded THCpSiNPs have been evaluated in vitro. Standard cell viability assays demonstrate that cells incubated with the NPs at a concentration of 0.1 mg/mL are 95% viable. In addition, chitosan coating significantly enhances the uptake of oligonucleotide-loaded THCpSiNPs across the cell membrane. Moreover, histopathological analysis of liver, kidney, spleen, and skin tissue collected from mice receiving NPs further demonstrates the biocompatible and non-inflammatory properties of the NPs as a gene delivery vehicle for intravenous and subcutaneous administration in vivo. Taken together, these results suggest that THCpSiNPs provide a versatile platform that could be used as efficient vehicles for the intracellular delivery of oligonucleotides for gene therapy.

关键词： nanoparticles porous silicon chitosan gene delivery

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