The Experts below are selected from a list of 186 Experts worldwide ranked by ideXlab platform
Nicholas T. Ventham - One of the best experts on this subject based on the ideXlab platform.
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Correction: Changes to Serum Sample Tube and Processing Methodology Does Not Cause Inter-Individual Variation in Automated Whole Serum N-Glycan Profiling in Health and Disease
PloS one, 2015Co-Authors: Nicholas T. Ventham, Richard A. Gardner, Nicholas A. Kennedy, Archana Shubhakar, Rahul Kalla, Elaine R. Nimmo, Daryl L. Fernandes, Jack Satsangi, Daniel SpencerAbstract:Introduction Serum N-glycans have been identified as putative biomarkers for numerous diseases. The impact of different serum Sample Tubes and processing methods on N-glycan analysis has received relatively little attention. This study aimed to determine the effect of different Sample Tubes and processing methods on the whole serum N-glycan profile in both health and disease. A secondary objective was to describe a robot automated N-glycan release, labeling and cleanup process for use in a biomarker discovery system. Methods 25 patients with active and quiescent inflammatory bowel disease and controls had three different serum Sample Tubes taken at the same draw. Two different processing methods were used for three types of Tube (with and without gel-separation medium). Samples were randomised and processed in a blinded fashion. Whole serum N-glycan release, 2-aminobenzamide labeling and cleanup was automated using a Hamilton Microlab STARlet Liquid Handling robot. Samples were analysed using a hydrophilic interaction liquid chromatography/ethylene bridged hybrid(BEH) column on an ultra-high performance liquid chromatography instrument. Data were analysed quantitatively by pairwise correlation and hierarchical clustering using the area under each chromatogram peak. Qualitatively, a blinded assessor attempted to match chromatograms to each individual. Results There was small intra-individual variation in serum N-glycan profiles from Samples collected using different Sample processing methods. Intra-individual correlation coefficients were between 0.99 and 1. Unsupervised hierarchical clustering and principal coordinate analyses accurately matched Samples from the same individual. Qualitative analysis demonstrated good chromatogram overlay and a blinded assessor was able to accurately match individuals based on chromatogram profile, regardless of disease status. Conclusions The three different serum Sample Tubes processed using the described methods cause minimal inter-individual variation in serum whole N-glycan profile when processed using an automated workstream. This has important implications for N-glycan biomarker discovery studies using different serum processing standard operating procedures.
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Changes to serum Sample Tube and processing methodology does not cause Intra-Individual [corrected] variation in automated whole serum N-glycan profiling in health and disease.
PloS one, 2015Co-Authors: Nicholas T. Ventham, Richard A. Gardner, Nicholas A. Kennedy, Archana Shubhakar, Rahul Kalla, Elaine R. Nimmo, Daryl L. Fernandes, Jack Satsangi, Daniel I R SpencerAbstract:Serum N-glycans have been identified as putative biomarkers for numerous diseases. The impact of different serum Sample Tubes and processing methods on N-glycan analysis has received relatively little attention. This study aimed to determine the effect of different Sample Tubes and processing methods on the whole serum N-glycan profile in both health and disease. A secondary objective was to describe a robot automated N-glycan release, labeling and cleanup process for use in a biomarker discovery system. 25 patients with active and quiescent inflammatory bowel disease and controls had three different serum Sample Tubes taken at the same draw. Two different processing methods were used for three types of Tube (with and without gel-separation medium). Samples were randomised and processed in a blinded fashion. Whole serum N-glycan release, 2-aminobenzamide labeling and cleanup was automated using a Hamilton Microlab STARlet Liquid Handling robot. Samples were analysed using a hydrophilic interaction liquid chromatography/ethylene bridged hybrid(BEH) column on an ultra-high performance liquid chromatography instrument. Data were analysed quantitatively by pairwise correlation and hierarchical clustering using the area under each chromatogram peak. Qualitatively, a blinded assessor attempted to match chromatograms to each individual. There was small intra-individual variation in serum N-glycan profiles from Samples collected using different Sample processing methods. Intra-individual correlation coefficients were between 0.99 and 1. Unsupervised hierarchical clustering and principal coordinate analyses accurately matched Samples from the same individual. Qualitative analysis demonstrated good chromatogram overlay and a blinded assessor was able to accurately match individuals based on chromatogram profile, regardless of disease status. The three different serum Sample Tubes processed using the described methods cause minimal inter-individual variation in serum whole N-glycan profile when processed using an automated workstream. This has important implications for N-glycan biomarker discovery studies using different serum processing standard operating procedures.
Sandeep Kumar Vashist - One of the best experts on this subject based on the ideXlab platform.
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evaluation of apparent non specific protein loss due to adsorption on Sample Tube surfaces and or altered immunogenicity
Analyst, 2011Co-Authors: Chandra K. Dixit, Sandeep Kumar Vashist, Brian D. Maccraith, Richard OkennedyAbstract:The non-specific loss of protein analytes can have a major effect on assay results particularly where the concentrations of such analytes are extremely low and the matrix is complex. This report assesses how the protein incubated in Sample Tubes may be lost due to adsorption. Use of proteins, such as bovine serum albumin (BSA), may be used to pre-treat Tubes to reduce such losses. However, such losses may also be associated with structural perturbations leading to changes in immunogenicity (as a result of alterations in specific epitope-related conformations). This can lead to erroneous results or lack of comparability with a range of methodologies such as the bicinchoninic protein assay and immunoassays or when surface plasmon resonance (SPR)-based approaches are used. A model system to evaluate these phenomena is proposed.
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Evaluation of apparent non-specific protein loss due to adsorption on Sample Tube surfaces and/or altered immunogenicity.
The Analyst, 2011Co-Authors: Chandra K. Dixit, Sandeep Kumar Vashist, Brian D. Maccraith, Richard O'kennedyAbstract:The non-specific loss of protein analytes can have a major effect on assay results particularly where the concentrations of such analytes are extremely low and the matrix is complex. This report assesses how the protein incubated in Sample Tubes may be lost due to adsorption. Use of proteins, such as bovine serum albumin (BSA), may be used to pre-treat Tubes to reduce such losses. However, such losses may also be associated with structural perturbations leading to changes in immunogenicity (as a result of alterations in specific epitope-related conformations). This can lead to erroneous results or lack of comparability with a range of methodologies such as the bicinchoninic protein assay and immunoassays or when surface plasmon resonance (SPR)-based approaches are used. A model system to evaluate these phenomena is proposed.
Chandra K. Dixit - One of the best experts on this subject based on the ideXlab platform.
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evaluation of apparent non specific protein loss due to adsorption on Sample Tube surfaces and or altered immunogenicity
Analyst, 2011Co-Authors: Chandra K. Dixit, Sandeep Kumar Vashist, Brian D. Maccraith, Richard OkennedyAbstract:The non-specific loss of protein analytes can have a major effect on assay results particularly where the concentrations of such analytes are extremely low and the matrix is complex. This report assesses how the protein incubated in Sample Tubes may be lost due to adsorption. Use of proteins, such as bovine serum albumin (BSA), may be used to pre-treat Tubes to reduce such losses. However, such losses may also be associated with structural perturbations leading to changes in immunogenicity (as a result of alterations in specific epitope-related conformations). This can lead to erroneous results or lack of comparability with a range of methodologies such as the bicinchoninic protein assay and immunoassays or when surface plasmon resonance (SPR)-based approaches are used. A model system to evaluate these phenomena is proposed.
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Evaluation of apparent non-specific protein loss due to adsorption on Sample Tube surfaces and/or altered immunogenicity.
The Analyst, 2011Co-Authors: Chandra K. Dixit, Sandeep Kumar Vashist, Brian D. Maccraith, Richard O'kennedyAbstract:The non-specific loss of protein analytes can have a major effect on assay results particularly where the concentrations of such analytes are extremely low and the matrix is complex. This report assesses how the protein incubated in Sample Tubes may be lost due to adsorption. Use of proteins, such as bovine serum albumin (BSA), may be used to pre-treat Tubes to reduce such losses. However, such losses may also be associated with structural perturbations leading to changes in immunogenicity (as a result of alterations in specific epitope-related conformations). This can lead to erroneous results or lack of comparability with a range of methodologies such as the bicinchoninic protein assay and immunoassays or when surface plasmon resonance (SPR)-based approaches are used. A model system to evaluate these phenomena is proposed.
Daniel I R Spencer - One of the best experts on this subject based on the ideXlab platform.
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Changes to serum Sample Tube and processing methodology does not cause Intra-Individual [corrected] variation in automated whole serum N-glycan profiling in health and disease.
PloS one, 2015Co-Authors: Nicholas T. Ventham, Richard A. Gardner, Nicholas A. Kennedy, Archana Shubhakar, Rahul Kalla, Elaine R. Nimmo, Daryl L. Fernandes, Jack Satsangi, Daniel I R SpencerAbstract:Serum N-glycans have been identified as putative biomarkers for numerous diseases. The impact of different serum Sample Tubes and processing methods on N-glycan analysis has received relatively little attention. This study aimed to determine the effect of different Sample Tubes and processing methods on the whole serum N-glycan profile in both health and disease. A secondary objective was to describe a robot automated N-glycan release, labeling and cleanup process for use in a biomarker discovery system. 25 patients with active and quiescent inflammatory bowel disease and controls had three different serum Sample Tubes taken at the same draw. Two different processing methods were used for three types of Tube (with and without gel-separation medium). Samples were randomised and processed in a blinded fashion. Whole serum N-glycan release, 2-aminobenzamide labeling and cleanup was automated using a Hamilton Microlab STARlet Liquid Handling robot. Samples were analysed using a hydrophilic interaction liquid chromatography/ethylene bridged hybrid(BEH) column on an ultra-high performance liquid chromatography instrument. Data were analysed quantitatively by pairwise correlation and hierarchical clustering using the area under each chromatogram peak. Qualitatively, a blinded assessor attempted to match chromatograms to each individual. There was small intra-individual variation in serum N-glycan profiles from Samples collected using different Sample processing methods. Intra-individual correlation coefficients were between 0.99 and 1. Unsupervised hierarchical clustering and principal coordinate analyses accurately matched Samples from the same individual. Qualitative analysis demonstrated good chromatogram overlay and a blinded assessor was able to accurately match individuals based on chromatogram profile, regardless of disease status. The three different serum Sample Tubes processed using the described methods cause minimal inter-individual variation in serum whole N-glycan profile when processed using an automated workstream. This has important implications for N-glycan biomarker discovery studies using different serum processing standard operating procedures.
Daniel Spencer - One of the best experts on this subject based on the ideXlab platform.
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Correction: Changes to Serum Sample Tube and Processing Methodology Does Not Cause Inter-Individual Variation in Automated Whole Serum N-Glycan Profiling in Health and Disease
PloS one, 2015Co-Authors: Nicholas T. Ventham, Richard A. Gardner, Nicholas A. Kennedy, Archana Shubhakar, Rahul Kalla, Elaine R. Nimmo, Daryl L. Fernandes, Jack Satsangi, Daniel SpencerAbstract:Introduction Serum N-glycans have been identified as putative biomarkers for numerous diseases. The impact of different serum Sample Tubes and processing methods on N-glycan analysis has received relatively little attention. This study aimed to determine the effect of different Sample Tubes and processing methods on the whole serum N-glycan profile in both health and disease. A secondary objective was to describe a robot automated N-glycan release, labeling and cleanup process for use in a biomarker discovery system. Methods 25 patients with active and quiescent inflammatory bowel disease and controls had three different serum Sample Tubes taken at the same draw. Two different processing methods were used for three types of Tube (with and without gel-separation medium). Samples were randomised and processed in a blinded fashion. Whole serum N-glycan release, 2-aminobenzamide labeling and cleanup was automated using a Hamilton Microlab STARlet Liquid Handling robot. Samples were analysed using a hydrophilic interaction liquid chromatography/ethylene bridged hybrid(BEH) column on an ultra-high performance liquid chromatography instrument. Data were analysed quantitatively by pairwise correlation and hierarchical clustering using the area under each chromatogram peak. Qualitatively, a blinded assessor attempted to match chromatograms to each individual. Results There was small intra-individual variation in serum N-glycan profiles from Samples collected using different Sample processing methods. Intra-individual correlation coefficients were between 0.99 and 1. Unsupervised hierarchical clustering and principal coordinate analyses accurately matched Samples from the same individual. Qualitative analysis demonstrated good chromatogram overlay and a blinded assessor was able to accurately match individuals based on chromatogram profile, regardless of disease status. Conclusions The three different serum Sample Tubes processed using the described methods cause minimal inter-individual variation in serum whole N-glycan profile when processed using an automated workstream. This has important implications for N-glycan biomarker discovery studies using different serum processing standard operating procedures.
