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S Blirup-Jensen

Publications and source records attributed to S Blirup-Jensen.

9 recordsLinked to original sources
PubMed2001

Protein standardization I: Protein purification procedure for the purification of human prealbumin, orosomucoid and transferrin as primary protein preparations.

A procedure for the purification of human prealbumin, orosomucoid and transferrin as primary protein preparations has been developed. The procedure describes in detail the chemicals, the fractionation equipment and the purification of the three proteins from the same starting material (a 90-donor plasma pool). The fractionation steps involve methods like salting out, various anion and cation exchange chromatographies, preparative electrophoresis, and finally, size chromatography. Only mild and highly reproducible fractionation methods are used in order to obtain high recoveries. All of these provide the best guarantee that no serious subfractionation has taken place. At the same time, high recoveries are a necessity for the pure protein to be representative of the same protein in vivo. The following recoveries were obtained: prealbumin 55%, orosomucoid 70% and transferrin 85%. The pure proteins are produced as liquid calibrators (primary protein preparations) dissolved in one electrolyte (0.1 mol/l KCl) and stored in sealed glass ampoules at -80 degrees C. These three pure proteins were used as primary reference preparations in the certification of the international reference preparation CRM 470.

Chromatography, Ion Exchange↗
PubMed2001

Protein standardization II: Dry mass determination procedure for the determination of the dry mass of a pure protein preparation.

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.

Blood Proteins↗
PubMed2001

Protein standardization III: Method optimization basic principles for quantitative determination of human serum proteins on automated instruments based on turbidimetry or nephelometry.

Quantitative protein determinations in routine laboratories are today most often carried out using automated instruments. However, slight variations in the assay principle, in the programming of the instrument or in the reagents may lead to different results. This has led to the prerequisite of method optimization and standardization. The basic principles of turbidimetry and nephelometry are discussed. The different reading principles are illustrated and investigated. Various problems are identified and a suggestion is made for an integrated, fast and convenient test system for the determination of a number of different proteins on the same instrument. An optimized test system for turbidimetry and nephelometry should comprise high-quality antibodies, calibrators, controls, and buffers and a protocol with detailed parameter settings in order to program the instrument correctly. A good user program takes full advantage of the optimal reading principles for the different instruments. This implies--for all suitable instruments--sample preincubation followed by real sample blanking, which automatically corrects for initial turbidity in the sample. Likewise it is recommended to measure the reagent blank, which represents any turbidity caused by the antibody itself. By correcting all signals with these two blank values the best possible signal is obtained for the specific analyte. An optimized test system should preferably offer a wide measuring range combined with a wide security range, which for the user means few re-runs and maximum security against antigen excess. A non-linear calibration curve based on six standards is obtained using a suitable mathematical fitting model, which normally is part of the instrument software.

Antigen-Antibody Reactions↗
PubMed2001

Protein standardization IV: Value transfer procedure for the assignment of serum protein values from a reference preparation to a target material.

A new approach for the assignment of values to serum proteins in a target material using a reference preparation has been developed. The procedure describes the general as well as the practical principles involved in the value assignment (with examples). Two models have been developed: 1) The direct value transfer between serum matrices and 2) the indirect value transfer from a pure protein preparation to a serum protein material. The necessary mathematical equations are developed and explained. The data reduction and statistical evaluation are discussed. The practical procedure (the transfer protocol) is based on six dilutions of the reference preparation assayed together with six dilutions of the target material. In this way imprecision is reduced and the proportionality of the two materials (i.e. the presence or absence of matrix effects) can be assessed directly by evaluating a single regression plot. If no matrix effects are found, the regression line will pass through zero with a slope equal to the ratio of the concentrations of the two materials. The transfer protocol is based on a multiple point value assignment obtained by several measurements a day repeated on several days, an important prerequisite being that all reconstitutions and dilutions are controlled by weighing.

Blood Proteins↗
PubMed1996

Recommendations for the selection and use of protocols for assignment of values to reference materials.

It is essential that testing of patient samples give values that are traceable to those in a recognized, authorizative reference material. In addition, samples used for laboratory proficiency testing must have values assigned from such a reference material if results are to be comparable among materials and laboratories. As a result, the assignment of values to secondary and tertiary reference materials, calibrants, controls, and proficiency samples should be performed as precisely as possible, within reasonable limits. The intent of this document is to give guidelines for assignment of values at three levels of transfer. 1) from primary to secondary reference, materials, such as international or national references; 2) from secondary to tertiary reference materials, such as manufacturers' in-house calibrants and controls; and 3) from tertiary reference materials, such as manufacturers' in-house calibrants and controls; and 3) from tertiary reference materials to working calibrants and controls. It is hoped that these guidelines will facilitate the selection and utilization of an appropriate value transfer protocol for each level of value assignment. Because of the wide variety and nature of analytes, however, the guidelines are intentionally broad and may require revision for specific analytes.

Calibration↗
PubMed1996

Consensus of a group of professional societies and diagnostic companies on guidelines for interim reference ranges for 14 proteins in serum based on the standardization against the IFCC/BCR/CAP Reference Material (CRM 470). International Federation of Clinical Chemistry. Community Bureau of Reference of the Commission of the European Communities. College of American Pathologists.

The release in 1993 of a new reference material for serum proteins, CRM 470/RPPHS 5 has given rise to a great improvement in the between-laboratory variability of serum protein measurements worldwide. Conversion to the new reference material results in significant changes in reference values for some proteins. The establishment of new reference ranges will take a considerable time, and in the interim several professional societies and diagnostic companies have agreed to use consensus reference ranges based on studies already undertaken.

Blood Proteins↗
PubMed1995

Characterization and quantification of plasma proteins excreted in faeces from healthy humans.

Faecal plasma protein loss was studied in 38 healthy adults. Using crossed immunoelectrophoresis and single radial immunodiffusion the most frequently found proteins were alpha 1-antitrypsin, IgA, alpha 1-antichymotrypsin (found in 97, 92, and 84% of subjects), prealbumin and IgM (both found in 55%). The major plasma proteins, albumin and IgG, were found in 37 and 13% of subjects, respectively, and in trace amounts only. alpha 2-macroglobulin could not be detected. There was no relation between the presence of proteins in faeces and their plasma concentration. When added to faeces, alpha 1-antitrypsin, alpha 1-antichymotrypsin, and prealbumin were resistant to incubation (37 degrees C, 48 h), whereas albumin, IgG, IgM, and IgA were rapidly degraded (within 8-24 h). Some IgA was bound to secretory component, indicating enteric secretion. alpha 2-macroglobulin was semi-resistant to degradation, but its passage to the intestinal lumen may have been prevented by its molecular size. In conclusion, resistance to degradation, enteric secretion, and low molecular weight are the primary factors which favour the excretion of plasma proteins in faeces. The technique used in this study allows further studies in patients with inflammatory changes and protein-losing enteropathy.

Adult↗
PubMed1994

New international reference preparation for proteins in human serum (RPPHS).

Quality-control surveys in recent years, in various parts of the world, have shown poor between-laboratory agreement for measurements of plasma proteins. Despite the existence of international reference materials distributed by the World Health Organization, standards produced by diagnostics manufacturers and professional organizations differ significantly in their ascribed values. The reasons for this are complex but include poor availability of the primary materials, confusion about their use, and the fact that their turbidity on reconstitution precludes their use in modern optical immunoassays. This unfortunate situation led to an important initiative to produce sufficient quantities of a widely available, optically clear secondary reference material for plasma proteins that could be used worldwide by manufacturers, professional organizations, and laboratories. Here we present an overview on how the laboratory community, including manufacturers, clinical laboratories, professional societies, and regulators, has reached what we consider is a successful conclusion to a difficult problem.

Blood Proteins↗