Protein Standardization II: Dry Mass Determination. Procedure for the Determination of the Dry Mass of a Pure Protein Preparation
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Søren Blirup-Jensen
Abstract
A procedure has been developed for the determination of the dry mass of a pure protein preparation dissolved in one electrolyte. The procedure not only renders the concentration (in g/l), but additionally gives the partial specific volume of the protein (in ml/g). The latter is an important parameter for the characterization of a specific protein. Furthermore, the importance of extensive dialysis against one electrolyte is discussed. For the drying of human serum proteins it is clearly shown that KCl is preferred to NaCl due to its stable temperature curve. By observing the parallel fluctuations in the weight of the empty vials during drying, an average correction factor is introduced, which greatly minimizes these changes. The assay principle is discussed from a mathematical as well as from a practical point of view. A detailed procedure is described and the final results for three primary pure proteins (prealbumin, orosomucoid and transferrin) are presented. Finally, important parameters such as the wavelength of maximum absorbance, the absorption coefficient and the specific refractive increment are discussed and values for the three proteins are presented. When these parameters have been established the future determination of concentration and the characterization of pure protein solutions are greatly facilitated. These procedures were important tools for ascribing mass values for prealbumin, orosomucoid and transferrin to the international reference preparation CRM 470.
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
Articles in the same Issue
- The Present State of Protein Analysis and Interpretation
- In Memoriam Carl-Bertil Laurell
- From Paper Electrophoresis to Computer-supported Interpretation of Capillary Electrophoresis – Clinical Plasma Protein Analysis in Malmö, Sweden
- The Pavia Approach to Clinical Protein Analysis
- How the Foundation for Blood Research (FBR) Has Managed Serum Protein Testing for New England Clinicians
- Evidence-Based Laboratory Interpretation System Built on a Large Collection of Case Records with Well-Defined Diagnoses
- A Knowledge-Based System to Aid with the Clinical Interpretation of Complex Serum Protein Data
- Markers of the Acute Phase Response in Cardiovascular Disease: An Update
- Protein Aggregation
- Protein Standardization I: Protein Purification. Procedure for the Purification of Human Prealbumin, Orosomucoid and Transferrin as Primary Protein Preparations
- Protein Standardization II: Dry Mass Determination. Procedure for the Determination of the Dry Mass of a Pure Protein Preparation
- Protein Standardization III: Method Optimization. Basic Principles for Quantitative Determination of Human Serum Proteins on Automated Instruments Based on Turbidimetry or Nephelometry
- Protein Standardization IV: Value Transfer. Procedure for the Assignment of Serum Protein Values from a Reference Preparation to a Target Material
- Effect of Certified Reference Material 470 (CRM 470) on National Quality Assurance Programs for Serum Proteins in Europe
- Commutability of Serum Protein Values: Persisting Bias among Manufacturers Using Values Assigned from the Certified Reference Material 470 (CRM 470) in the United States
- The Existing Interim Consensus Reference Ranges and the Future Approach
- Using Multiples of the Median to Normalize Serum Protein Measurements
- Strategy for Determining Racial and Environmental Similarities and Differences for Plasma Proteins
- Standardization of Immunoassay for CRM-Related Proteins in Japan: From Evaluating CRM 470 to Setting Reference Intervals
- Soluble Transferrin Receptor (sTfR): Biological Variations and Reference Limits
- High Sensitivity C-Reactive Protein (CRP) Reference Intervals in the Elderly
- High Sensitivity Immunoassays for C-Reactive Protein: Promises and Pitfalls
- Meetings and Awards
Articles in the same Issue
- The Present State of Protein Analysis and Interpretation
- In Memoriam Carl-Bertil Laurell
- From Paper Electrophoresis to Computer-supported Interpretation of Capillary Electrophoresis – Clinical Plasma Protein Analysis in Malmö, Sweden
- The Pavia Approach to Clinical Protein Analysis
- How the Foundation for Blood Research (FBR) Has Managed Serum Protein Testing for New England Clinicians
- Evidence-Based Laboratory Interpretation System Built on a Large Collection of Case Records with Well-Defined Diagnoses
- A Knowledge-Based System to Aid with the Clinical Interpretation of Complex Serum Protein Data
- Markers of the Acute Phase Response in Cardiovascular Disease: An Update
- Protein Aggregation
- Protein Standardization I: Protein Purification. Procedure for the Purification of Human Prealbumin, Orosomucoid and Transferrin as Primary Protein Preparations
- Protein Standardization II: Dry Mass Determination. Procedure for the Determination of the Dry Mass of a Pure Protein Preparation
- Protein Standardization III: Method Optimization. Basic Principles for Quantitative Determination of Human Serum Proteins on Automated Instruments Based on Turbidimetry or Nephelometry
- Protein Standardization IV: Value Transfer. Procedure for the Assignment of Serum Protein Values from a Reference Preparation to a Target Material
- Effect of Certified Reference Material 470 (CRM 470) on National Quality Assurance Programs for Serum Proteins in Europe
- Commutability of Serum Protein Values: Persisting Bias among Manufacturers Using Values Assigned from the Certified Reference Material 470 (CRM 470) in the United States
- The Existing Interim Consensus Reference Ranges and the Future Approach
- Using Multiples of the Median to Normalize Serum Protein Measurements
- Strategy for Determining Racial and Environmental Similarities and Differences for Plasma Proteins
- Standardization of Immunoassay for CRM-Related Proteins in Japan: From Evaluating CRM 470 to Setting Reference Intervals
- Soluble Transferrin Receptor (sTfR): Biological Variations and Reference Limits
- High Sensitivity C-Reactive Protein (CRP) Reference Intervals in the Elderly
- High Sensitivity Immunoassays for C-Reactive Protein: Promises and Pitfalls
- Meetings and Awards
