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排序方式: 共有177条查询结果,搜索用时 31 毫秒
41.
Eric E. Niederkofler David A. Phillips Bryan Krastins Vathany Kulasingam Urban A. Kiernan Kemmons A. Tubbs Scott M. Peterman Amol Prakash Eleftherios P. Diamandis Mary F. Lopez Dobrin Nedelkov 《PloS one》2013,8(11)
Insulin-like growth factor 1 (IGF1) is an important biomarker of human growth disorders that is routinely analyzed in clinical laboratories. Mass spectrometry-based workflows offer a viable alternative to standard IGF1 immunoassays, which utilize various pre-analytical preparation strategies. In this work we developed an assay that incorporates a novel sample preparation method for dissociating IGF1 from its binding proteins. The workflow also includes an immunoaffinity step using antibody-derivatized pipette tips, followed by elution, trypsin digestion, and LC-MS/MS separation and detection of the signature peptides in a selected reaction monitoring (SRM) mode. The resulting quantitative mass spectrometric immunoassay (MSIA) exhibited good linearity in the range of 1 to 1,500 ng/mL IGF1, intra- and inter-assay precision with CVs of less than 10%, and lowest limits of detection of 1 ng/mL. The linearity and recovery characteristics of the assay were also established, and the new method compared to a commercially available immunoassay using a large cohort of human serum samples. The IGF1 SRM MSIA is well suited for use in clinical laboratories. 相似文献
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Nedelkov D Kiernan UA Niederkofler EE Tubbs KA Nelson RW 《Molecular & cellular proteomics : MCP》2006,5(10):1811-1818
This review outlines the concept of population proteomics and its implication in the discovery and validation of cancer-specific protein modulations. Population proteomics is an applied subdiscipline of proteomics engaging in the investigation of human proteins across and within populations to define and better understand protein diversity. Population proteomics focuses on interrogation of specific proteins from large number of individuals, utilizing top-down, targeted affinity mass spectrometry approaches to probe protein modifications. Deglycosylation, sequence truncations, side-chain residue modifications, and other modifications have been reported for myriad of proteins, yet little is know about their incidence rate in the general population. Such information can be gathered via population proteomics and would greatly aid the biomarker discovery efforts. Discovery of novel protein modifications is also expected from such large scale population proteomics, expanding the protein knowledge database. In regard to cancer protein biomarkers, their validation via population proteomics-based approaches is advantageous as mass spectrometry detection is used both in the discovery and validation process, which is essential for the detection of those structurally modified protein biomarkers. 相似文献
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Background
Understanding evolutionary processes that drive genome reduction requires determining the tempo (rate) and the mode (size and types of deletions) of gene losses. In this study, we analysed five endosymbiotic genome sequences of the gamma-proteobacteria (three different Buchnera aphidicola strains, Wigglesworthia glossinidia, Blochmannia floridanus) to test if gene loss could be driven by the selective importance of genes. We used a parsimony method to reconstruct a minimal ancestral genome of insect endosymbionts and quantified gene loss along the branches of the phylogenetic tree. To evaluate the selective or functional importance of genes, we used a parameter that measures the level of adaptive codon bias in E. coli (i.e. codon adaptive index, or CAI), and also estimates of evolutionary rates (Ka) between pairs of orthologs either in free-living bacteria or in pairs of symbionts. 相似文献45.
Insulin like growth factor (IGF)-1 and IGF-2 were assayed from human plasma via biomolecular interaction analysis mass spectrometry, utilizing antibodies as ligands for affinity retrieval. Detection of both targeted and non-targeted IGFs in the mass spectra indicated possible protein complex retrieval by the individual antibodies. A series of control experiments eliminated the possibility of analyte cross-walking between flow cells, significant antibodies cross-reactivity, and direct IGF interactions. To disrupt the putative protein complex and release its constituent proteins, plasma samples were treated with detergents. An SDS-treated plasma yielded IGF signals in a different ratio than the one observed in the mass spectra from the non-treated plasma, suggesting disruption of the protein complex, and its retrieval from non-treated plasma. Novel truncated IGF-2 variant, missing its N-terminal Alanine, was detected in all mass spectra. 相似文献
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Fragments of adult renal tissue were maintained as organ cultures for 7 days in HEPES-buffered medium 199, supplemented with 10% calf serum and antibiotics. Addition of aprotinin (10,000 kallikrein inhibitor units per ml) to the medium resulted in improved survival of the cells of the glomeruli and tubules and preserved the integrity of the glomerular, capsular and tubular basement membranes. Optimal results were obtained when aprotinin was present throughout the period of culturing. Inclusion of aprotinin in the medium for only the first 3 days in vitro slightly increased the numbers of surviving glomerular and tubular cells, but also promoted the growth of connective tissue in the explants and was detrimental to the basement membranes of the tubules. It is suggested that both the antiproteolytic and the carbohydrate-binding properties of aprotinin are involved in the mediation of the observed effects. 相似文献
48.
Galbeta1-3GalNAc (T-disaccharide) and related molecules were assayed to
describe the structural requirements of carbohydrates to bind Agaricus
bisporus lectin (ABL). Results provide insight into the most relevant
regions of T-disaccharide involved in the binding of ABL. It was found that
monosaccharides bind ABL weakly indicating a more extended
carbohydrate-binding site as compared to those involvedin the T-
disaccharide specific lectins such as jacalin and peanut agglutinin.
Lacto-N-biose (Galbeta1-3GlcNAc) unlike T-disaccharide, is unable to
inhibit the ABL interaction, thus showing the great importance of the
position of the axial C-4 hydroxyl group of GalNAc in T-disaccharide. This
finding could explain the inhibitory ability of Galbeta1-6GlcNAc and
lactose because C-4 and C-3 hydroxyl groups of reducing Glc, respectively,
occupy a similar position as reported by conformational analysis. From the
comparison of different glycolipids bearing terminal T-disaccharide bound
to different linkages, it can be seen than ABL binding is even more
impaired by an adjacent C-6 residual position than by the anomeric
influence of T-disaccharide. Furthermore, the addition of beta-GlcNAc to
the terminal T-disaccharide in C-3 position of Gal does not affect the ABL
binding whereas if an anionic group such as glucuronic acid is added to
C-3, the binding is partially affected. These findings demonstrate that ABL
holds a particular binding nature different from that of other
T-disaccharide specific lectins.
相似文献
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