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- Ohlson, Sten, et al.
(författare)
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High-Performance Liquid Affinity Chromatography: Rapid Immunoanalysis of Transferrin in Serum
- 1988
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Ingår i: Clinical Chemistry. - 0009-9147 .- 1530-8561. ; 34:10, s. 2039-2043
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Tidskriftsartikel (refereegranskat)abstract
- We describe a new method for quantitatively measuring substances ofclinical interest by high-performance liquid affinity chromatography(HPLAC). As a model system we selected analysis for transferrin in humanserum with immobilized antibodies in a high-performance liquidchromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or 10 x0.5 cm) were used for in situ coupling of polyclonal antibodies totransferrin. The amount of transferrin eluted was determined by integratingthe eluted peak at 280 nm. The whole analytical procedure-- includinginjection of sample, washing, elution, and analysis of data-- takes only 7min. We characterized the HPLAC system for analysis of transferrin inseveral ways: intra-assay CV approximately 3%; inter- assay CV 2-9%; linearresponse up to 1 mg/mL column volume; detection limit approximately 3micrograms; analytical recovery 98% +/- 2%; purity of eluted sample greaterthan 95% (SDS-PAGE). The HPLAC method was compared with "rocket"immunoelectrophoresis, a commonly used method of analysis for transferrin,and there was excellent correlation between the two methods (r = 0.96, n =60). Benefits of this HPLAC technique include high precision, rapidanalysis, and simplified sample handling.
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| 2. |
- Abrahamsson, Per-Anders, et al.
(författare)
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Radioimmunoassay of beta-microseminoprotein, a prostatic-secreted protein present in sera of both men and women
- 1989
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Ingår i: Clinical Chemistry. - : American Association for Clinical Chemistry. - 0009-9147. ; 35:7, s. 1497-1503
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Tidskriftsartikel (refereegranskat)abstract
- We describe a simple radioimmunoassay of beta-microseminoprotein, one of the three most abundant secretory proteins of the prostate gland. The detection limit of the assay is 1 microgram/L, and its precision, expressed as the total coefficient of variation, is less than 10% for values between 10 and 150 micrograms/L. Using this assay, we found that beta-microseminoprotein immunoreactivity was present in sera from both sexes at about the same concentration. The protein detected had the same molecular size on gel chromatography as the protein isolated from seminal plasma, and dilution curves for the sera paralleled that for the pure protein. The findings suggest that beta-microseminoprotein is present in serum of healthy subjects of both sexes and that it originates in tissue other than the prostate gland. The range of the serum concentration was 0-10.6 micrograms/L (median 4.1) for 51 healthy adult women and 1.1-14.7 micrograms/L (median 6.2) for 35 healthy adult men not older than 40 years. In males with prostatic cancer the concentration in serum was highly variable and often greatly increased. The concentration of beta-microseminoprotein was correlated with that of creatinine in serum, suggesting that the protein is eliminated--at least partly--from the circulation by glomerular filtration. Little of the protein was present in the urine of women. In urine from men the concentration was high and variable, probably because of local contribution from the prostate gland to the urethral urine.
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