We are analyzing https://link.springer.com/article/10.1007/s12033-014-9804-7.

Title:
Comparison of Colorimetric Assays with Quantitative Amino Acid Analysis for Protein Quantification of Generalized Modules for Membrane Antigens (GMMA) | Molecular Biotechnology
Description:
Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.
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Keywords {🔍}

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Topics {✒️}

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Schema {🗺️}

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         headline:Comparison of Colorimetric Assays with Quantitative Amino Acid Analysis for Protein Quantification of Generalized Modules for Membrane Antigens (GMMA)
         description:Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.
         datePublished:2014-09-16T00:00:00Z
         dateModified:2014-09-16T00:00:00Z
         pageStart:84
         pageEnd:93
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            Protein quantification
            Generalized Modules for Membrane Antigens (GMMA)
            Outer Membrane Vesicles (OMV)
            Quantitative amino acid analysis
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            Non-Interfering assay
            Biotechnology
            Biochemistry
            general
            Cell Biology
            Protein Science
            Biological Techniques
            Human Genetics
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ScholarlyArticle:
      headline:Comparison of Colorimetric Assays with Quantitative Amino Acid Analysis for Protein Quantification of Generalized Modules for Membrane Antigens (GMMA)
      description:Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.
      datePublished:2014-09-16T00:00:00Z
      dateModified:2014-09-16T00:00:00Z
      pageStart:84
      pageEnd:93
      sameAs:https://doi.org/10.1007/s12033-014-9804-7
      keywords:
         Protein quantification
         Generalized Modules for Membrane Antigens (GMMA)
         Outer Membrane Vesicles (OMV)
         Quantitative amino acid analysis
         Colorimetric protein assay
         Lowry assay
         Bradford assay
         Non-Interfering assay
         Biotechnology
         Biochemistry
         general
         Cell Biology
         Protein Science
         Biological Techniques
         Human Genetics
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            1073-6085
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                     name:Novartis Vaccines Institute for Global Health, Siena, Italy
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                  name:Novartis Vaccines Institute for Global Health
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            name:Novartis Vaccines Institute for Global Health
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            name:Novartis Vaccines Institute for Global Health
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               name:Novartis Vaccines Institute for Global Health, Siena, Italy
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            address:
               name:Novartis Vaccines Institute for Global Health, Siena, Italy
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      name:Calman A. MacLennan
      affiliation:
            name:Novartis Vaccines Institute for Global Health
            address:
               name:Novartis Vaccines Institute for Global Health, Siena, Italy
               type:PostalAddress
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            name:Novartis Vaccines Institute for Global Health
            address:
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      name:Novartis Vaccines Institute for Global Health, Siena, Italy
      name:Novartis Vaccines Institute for Global Health, Siena, Italy

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