共查询到14条相似文献,搜索用时 0 毫秒
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Among the gene therapy vectors developed to date, lentiviral vectors persist in the host and are therefore best suited for long-term gene transfer and gene-replacement therapies. Human immunodeficiency virus (HIV)-1-based lentiviral vector products are currently produced by transient transfection, filtration and ultracentrifugation, and the quality of the resultant vector product is variable. We reason that by identifying host proteins co-produced with viral vectors we may better understand the mechanism of viral vector production, and improve the safety and quality of the resultant products. Our LC-MS/MS studies identified both viral vector proteins and host proteins, including nuclear proteins, elongation factors and chaperone and heat shock proteins (HSP), and confirmed the presence of known HIV-incorporated proteins, e.g. elongation factor-1α, HSP70 and Histone 2A, demonstrating the capability and viability of LC-MS/MS methods for the proteomic analysis of highly complex samples. Evaluation of the functions of the identified components is in progress to understand their implication for product quality and safety. These studies support the development of improved production and characterisation methods and advance the clinical application of lentiviral vector-based products. 相似文献
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High throughput quantification of apolipoproteins A‐I and B‐100 by isotope dilution MS targeting fast trypsin releasable peptides without reduction and alkylation 下载免费PDF全文
Bryan A Parks David M Schieltz Michael L Andrews Michael S Gardner Jon C Rees Christopher A Toth Jeffrey I Jones Lisa G McWilliams Zsuzsanna Kuklenyik James L Pirkle John R Barr 《Proteomics. Clinical applications》2017,11(7-8)
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The most complete proteome of human lenses has been compiled using 2-D LC-MS/MS analysis of foetal, aged normal and advanced nuclear cataract lenses. A total of 231 proteins were identified across all lens groups, including 112 proteins that have not been reported previously. Proteins were grouped according to their PANTHER molecular function classification in order to facilitate comparisons. Previously unreported N-terminal acetylation was detected in a number of proteins, with the majority being associated with the prior removal of a methionine residue. This pattern of proteolysis may indicate that methionine aminopeptidase activity is present in human lenses. Acetylation is likely to aid in the stability of proteins that are present in the lens for many decades. Protein sequences were also used to interrogate the three human lens cDNA libraries publicly available. Surprisingly, 84 proteins we identified were not present in the cDNA libraries. 相似文献
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Boateng J Kay R Lancashire L Brown P Velloso C Bouloux P Teale P Roberts J Rees R Ball G Harridge S Goldspink G Creaser C 《Proteomics. Clinical applications》2009,3(8):912-922
An integrated MS-based proteomic approach is described that combines MALDI-MS and LC-MS with artificial neural networks for the identification of protein and peptide biomarkers associated with recombinant human growth hormone (rhGH) administration. Serum from exercised males administered with rhGH or placebo was analysed using ELISA to determine insulin-like growth factor-I concentrations. Diluted serum from rhGH- and placebo-treated subjects was analysed for protein biomarkers by MALDI-MS, whereas LC-MS was used to analyse tryptically digested ACN-depleted serum extracts for peptide biomarkers. Ion intensities and m/z values were used as inputs to artificial neural networks to classify samples into rhGH- and placebo-treated groups. Six protein ions (MALDI-MS) correctly classified 96% of samples into their respective groups, with a sensitivity of 91% (20 of 22 rhGH treated) and specificity of 100% (24 of 24 controls). Six peptide ions (LC-MS) were also identified and correctly classified 93% of samples with a sensitivity of 90% (19 of 21 rhGH treated) and a specificity of 95% (20 of 21 controls). The peptide biomarker ion with the highest significance was sequenced using LC-MS/MS and database searching and found to be associated with leucine-rich α-2-glycoprotein. 相似文献
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O'Riordan A Johnston O McMorrow T Wynne K Maguire P Hegarty JE McCormick A Watson AJ Cagney G Gallagher WM Ryan MP 《Proteomics. Clinical applications》2008,2(9):1338-1348
Chronic kidney disease (CKD) is a common complication post‐orthotopic liver transplantation (OLT). Development of CKD is detected by monitoring serum urea and creatinine, however disease can occasionally be at an advanced stage before they become abnormal. Therefore, more accurate parameters are required. In order to identify novel biomarkers of CKD, serum was obtained from 47 OLT recipients with CKD (glomerular filtration rate <60 mL/min) and 23 with normal renal function (glomerular filtration rate >90 mL/min). Using the proteomic technique SELDI‐TOF‐MS, three protein biomarkers (55.6 kDa, 9.5 kDa and 11.4 kDa) were identified that, together, could stratify patients into cases or controls with a sensitivity and specificity of 93.6 and 91.3%, respectively. The area under the curve was 0.94. The primary splitter of the groups at 55.6 kDa was an alternative version of a molecule at 27.8 kDa, which was subsequently identified by 1‐D SDS‐PAGE and LC‐ESI‐MS/MS to be Apolipoprotein AI. Protein expression was shown to be reduced in CKD, by both ELISA (p = 0.057) and Western blot analysis (p = 0.003). Apolipoprotein AI is a novel, accurate marker of CKD post‐OLT. It does require further validation in a large, more diverse patient population but could potentially improve detection of CKD. 相似文献
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Pancreatic cancer is a highly lethal disease that is difficult to diagnose at early stage and even more difficult to cure. SW1990 and PANC-1 represent the two cancer cell lines, which are both derived from pancreatic duct, but at different cell differentiation stages. In this study, we applied the iTRAQ-labeling technology and 2-D strong cation exchange/reversed phase liquid chromatography – LC-MS/MS) to profile the secreted proteins of SW1990 and PANC-1 cells in a conditioned cell culture medium. A total of 401 proteins were identified by MS/MS and protein database searching, the percentages of these proteins predicted in the categories of plasma membrane, intracellular and secreted proteins were 29.2, 32.7 and 38.2%, respectively. Fifty six proteins were identified with unknown functions and 19 proteins were quantified with significant level changes between the two cancer cell lines under the specific cell condition with 12 proteins being up-regulated (>1.3-fold change) in PANC-1 (e.g. FLJ31222 protein, 97 kDa protein, type IV collagenase precursor, 38 kDa protein and centaurin) and seven proteins being up-regulated in SW1990 (e.g. fibroblast growth factor receptor substrate 2, putative p150, hypothetical protein LOC 654463 and LOC 55701). The proteins with significant level changes may provide a baseline to investigate mechanisms underlying the differentiation of two cell lines and can be further screened for better protein biomarkers in pancreatic cancer. 相似文献
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Identification of potential breast cancer markers in nipple discharge by protein profile analysis using two‐dimensional nano‐liquid chromatography/nanoelectrospray ionization‐mass spectrometry 下载免费PDF全文
Sadamu Kurono Yuka Kaneko Nariaki Matsuura Haruki Oishi Shinzaburo Noguchi Seung Jin Kim Yasuhiro Tamaki Takao Aikawa Yoshikazu Kotsuma Hideo Inaji Shuji Matsuura 《Proteomics. Clinical applications》2016,10(5):605-613
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Harada T Kuramitsu Y Makino A Fujimoto M Iizuka N Hoshii Y Takashima M Tamesa M Nishimura T Takeda S Abe T Yoshino S Oka M Nakamura K 《Proteomics. Clinical applications》2007,1(2):215-223
To identify proteins associated with esophageal carcinogenesis, we performed protein profiling of 16 esophageal squamous cell carcinomas (ESCCs) and paired noncancerous tissues by 2-DE and MS/MS. In cancerous tissues, three spots showed significant up-regulation in the amount of protein, while eight spots were significantly down-regulated. The identities of the spots were determined by PMF with LC-MS/MS and were confirmed by immunoblotting. The up-regulated proteins were tropomyosin alpha 4 chain, transgelin, and pyruvate kinase. The down-regulated proteins were serum albumin precursor, isoforms of annexin A1, tropomyosin beta chain, 14-3-3 protein sigma, and isoforms of serotransferrin precursor. In all 16 cases, up-regulation of the tropomyosin alpha 4 chain was confirmed by immunoblotting. Localization of the tropomyosin alpha 4 chain in ESCC cells and adjacent fibroblasts was confirmed by immunohistochemistry. 相似文献
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Quantification by nano liquid chromatography parallel reaction monitoring mass spectrometry of human apolipoprotein A‐I,apolipoprotein B,and hemoglobin A1c in dried blood spots 下载免费PDF全文
Clark M. Henderson James G. Bollinger Jessica O. Becker Jennifer M. Wallace Thomas J. Laha Michael J. MacCoss Andrew N. Hoofnagle 《Proteomics. Clinical applications》2017,11(7-8)