Cleanascite™ for lipid removal and identification of biomarkers from fish bile samples containing metalloproteinases

MONMOUTH JUNCTION, NJ., July 15, 2012 -- Fish is ideal for assessing water-borne or sediment-deposited toxins, provides knowledge of environmental contamination due to toxicants and therefore comparing fish proteomes to analyze proteome expression under complex field situations could provide knowledge of gene products, metabolites, or proteins for further studies. Authors Rachel Hauser-Davis, Lima AA, Ziolli RL, Campos RC cites Cleanascite™ as an ideal lipid clarification reagent during sample preparation of fish bile containing matrix metalloproteinases (MMPs). The paper titled, "First-time report of metalloproteinases in fish bile and their potential as bioindicators regarding environmental contamination" is published in the journal Aquatic Toxicology.2012; 110-111:99-106. Scientists identified matrix metalloproteinases in the bile of mullets (Mugil liza) and tilapias (Tilapia rendalli) which required clarification and purification studies prior to performing SDS-PAGE and zymography analysis. Lipid removal was performed using the delipidizer Cleanascite™, which is a non-ionic lipid adsorbent, used to precipitate lipid fat droplets, cell debris and mucinous impurities. It is ideal for clarifying ascites, serum, cell & tissue culture, bile and organ homogenates. Metalloproteins in fish could be used as biomarkers of environmental exposure for fish and have utility as biomarkers for environmental contamination (López-Barea and Gómez-Ariza 2006 ). Quantifying total metal levels in fish organs, and directly relating them to environmental contamination provides information about toxic exposure levels (Moldovan et al. 2004 ).Total metal levels in fish are influenced by the amount of metals present in the form of metal-bound proteins and/or on-site contamination. In general, scientist describe Cleanascite™ for lipid clarification from gallbladder bile samples of 2 fish species, mullet (Mugil liza) and tilapias (Tilapia rendalli) containing substantial matrix metalloproteinases (MMPs). Extensive purification studies were conducted in order to obtain workable samples for SDS-PAGE and zymography analysis. Cleanascite™ is used in this experiment for lipid removal. Cleanascite™ is a solid-phase, non-ionic adsorbent supplied as a suspension in saline, ready for use. Simply add, centrifuge and/or filter. The clarified supernatant is ready for subsequent downstream processing or analysis. In this study scientists also conducted numerous inhibitor studies in which MMPs were inhibited. Differences in proteinase expression were observed in fish from a contaminated and reference site. Metalloproteinases in fish bile are routinely analyzed by mass spectrometry approaches such as inductively coupled plasma MS (ICP-MS), electrospray MS (ESI-MS), and matrix-assisted laser desorption ionization MS (MALDI-MS) alongside prior 2D-PAGE protein separation to rapidly and precisely identify gene products, metabolites, or proteins. About Biotech Support Group LLC Biotech Support Group LLC is a leading provider of genomics and proteomics sample preparation products and enrichment reagent kits as well as integrated biotechnology services for life sciences research, biomarker and drug discovery. Based in New Jersey, it's principal products include: AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, NuGel™ for passivated silica-based affinity chromatography, HemoVoid™ Hemoglobin Depletion From Erythrocytes, HemogloBind™ Hemoglobin Depletion From Hemolyzed Serum/Plasma and ProCipitate™ & ProPrep™ for nucleic acid isolation. Currently, Biotech Support Group LLC and ProFACT Proteomics Inc., are collaborating on the development of a proteomics platform used in functional profiling for proteomic analysis and a separations method for generating sub-proteomes used in biomarker and functional proteomic prospecting. For more information, go to CONTACT: Ravish Amin Biotech Support Group 1 Deer Park Drive, Suite M, Monmouth Junction, NJ 08852, USA 732-274-2866 [email protected] References for Cleanascite™ Hauser-Davis RA, Lima AA, Ziolli RL, Campos RC.First-time report of metalloproteinases in fish bile and their potential as bioindicators regarding environmental contamination. Aquatic Toxicology.2012;110-111:99-106 Farina A, Dumonceau JM, Frossard JL. Proteomic Analysis of Human Bile from Malignant Biliary Stenosis Induced by Pancreatic Cancer Journal of Proteome Research.2009; 8(1):159-69 Guerrier L, Claverol S, Finzi L et al. Contribution of solid-phase hexapeptide ligand libraries to the repertoire of human bile proteins. Journal of Chromatography.2007;1176(1-2):192-205 Chen Bo, Zheng Jian-wei, Wang Jian-ming, et al. Establishment and preliminary analysis of a 2-D human biliary map Chinese Journal of Hepatobiliary Surgery.2007 Chen B, Dong JQ, Chen YJ et al Two-dimensional electrophoresis for comparative proteomic analysis of human bile. Hepatobiliary & pancreatic diseases international.2007 Aug;6(4):402-6 Guerrier L, Claverol S, Finzi L et al Contribution of solid-phase hexapeptide ligand libraries to the repertoire of human bile proteins.Journal of Chromatography A.2007;1176(1-2):192-205 Kristiansen TZ, Bunkenborg J, Gronborg M et al A Proteomic Analysis of Human Bile Molecular and Cellular Proteomics.2004;3:715-728 Suggested References Proteomic analyses indicate induction of hepatic carbonyl reductase/20 beta-hydroxysteroid dehydrogenase B in rainbow trout exposed to sewage effluent. Ecotoxicology and environmental safety.1977; 68:33-39 Bode W, Maskos K. Structural basis of the matrix metalloproteinases and their physiological inhibitors, the tissue inhibitors of metalloproteinases. Biological Chemistry.2003; 384:863-872 Dowling VA, Sheehan D. Proteomics as a route to identification of toxicity targets in environmental toxicology. Proteomics. 2006; 6:5597-5604 Gonzalez-Fernandez M, Garcia-Barrera T, Arias-Borrego A, Jurado J, Pueyo C, Lopez-Barea J, Gomez-Ariza JL. Integrated application of transcriptomics, proteomics, and metallomics in environmental studies. Pure Applied Chemistry. 2008; 80:2609-2626 Egil Jellum, A. K. Thorsrud, F. W. Karasek. Two-dimensional electrophoresis for determining toxicity of environmental substances. Analytical chemistry.1983; 55(14):2340-2344 López-Barea J, Gómez-Ariza JL. Environmental proteomics and metallomics.Proteomics.2006; 6:S51-S62 Tay TL, Lin QS, Seow TK, Tan KH, Hew CL, Gong ZY. Proteomic analysis of protein profiles during early development of the zebrafi sh, Danio rerio.Proteomics.2006; 6:3176-3188 Shapiro SD. Diverse roles of macrophage matrix metalloproteinases in tissue destruction and tumor growth. Thrombosis and haemostasis. 1999;82(2):846-9 Wang JS, Wei YH, Wang DZ, Chan LL, Dai JY. Proteomic study of the effects of complex environmental stresses in the livers of goldfish (Carassius auratus) that inhabit Gaobeidian Lake in Beijing, China. 2008.Ecotoxicology 17:213-220

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