Abstract:
:We describe a new method for quantitatively measuring substances of clinical interest by high-performance liquid affinity chromatography (HPLAC). As a model system we selected analysis for transferrin in human serum with immobilized antibodies in a high-performance liquid chromatographic system. SelectiSpher-10 Activated Tresyl columns (5 or 10 x 0.5 cm) were used for in situ coupling of polyclonal antibodies to transferrin. The amount of transferrin eluted was determined by integrating the eluted peak at 280 nm. The whole analytical procedure--including injection of sample, washing, elution, and analysis of data--takes only 7 min. We characterized the HPLAC system for analysis of transferrin in several ways: intra-assay CV approximately 3%; inter-assay CV 2-9%; linear response up to 1 mg/mL column volume; detection limit approximately 3 micrograms; analytical recovery 98% +/- 2%; purity of eluted sample greater than 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, rapid analysis, and simplified sample handling.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Ohlson S,Gudmundsson BM,Wikström P,Larsson POsubject
Has Abstractpub_date
1988-10-01 00:00:00pages
2039-43issue
10eissn
0009-9147issn
1530-8561journal_volume
34pub_type
杂志文章abstract::We have developed a fully enzymatic method to measure 1,5 anhydro-D-glucitol (1,5-AG) in serum through use of pyranose oxidase (PROD: EC 1.1.3.10), glucokinase (EC 2.7.1.2), and an ATP-regenerating system. In a previous report (Clin Chem 1989;35:2039-43) the glucose interfering with the measurement of 1,5-AG was remov...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-11-01 00:00:00
abstract::The isoenzymes of lactate dehydrogenase (EC 1.1.1.27) were separated with excellent resolution in less than 1 h by high-pressure anion-exchange chromatography. These isoenzymes were determined in some tissue extracts and high-activity serum samples by this technique. ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-04-01 00:00:00
abstract::To secure a successful product in today's era of cost containment, manufacturers and developers of new in vitro diagnostic technologies must consider payment issues--coverage and reimbursement by healthcare insurers--as critical market forces that must be addressed in the business plan. Similar to the regulatory hurdl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1996-09-01 00:00:00
abstract::Consecutive values for serum osmolality measured in a patient exceeded (548 mOsm) the previously reported pathological range and, indeed, exceeded that generally considered compatible with life. The patient was receiving large amounts of intravenous dimethyl sulfoxide as part of an experimental protocol to control int...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-11-01 00:00:00
abstract::It has been estimated that approximately 37% of the US population judged to be at high risk for developing coronary artery disease (CAD), based on the National Cholesterol Education Program guidelines, have increased plasma lipoprotein(a) [Lp(a)], whereas Lp(a) is increased in only 14% of those judged to be at low ris...
journal_title:Clinical chemistry
pub_type:
doi:10.1373/clinchem.2003.023689
更新日期:2003-11-01 00:00:00
abstract::We compared results obtained with a commercial immunoradiometric assay kit for human thyrotropin in which monoclonal antibodies are used (Tandem -R TSH (ONE STEP) ImmunoRadioMetricAssay; Hybritech Inc.) with those obtained with two commercial radioimmunoassay methods: GAMMA-DAB [125I]HS-hTSH RIA (Clinical Assays) and ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-06-01 00:00:00
abstract::Cardiac troponin I (cTnI) is a regulatory protein unique to myocardium. We used a cardiospecific 30-min ELISA to measure cTnI in EDTA-plasma samples serially drawn from 28 patients before and after coronary artery bypass grafting (CABG)--26 elective and 2 salvage cases. The cTnI increase in 22 of the elective CABG pat...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1994-11-01 00:00:00
abstract::A method is described for separating antibody-bound from free fractions in ligand assays by column affinity chromatography, and its application to radioimmunoassay of choriomammotropin. In the method, 70 x 10 mm (i.d.) polypropylene columns containing about 150 mg of immunosorbent (goat anti-rabbit gamma-globulins cov...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1981-06-01 00:00:00
abstract:BACKGROUND:We previously reported a family in which the propositus and both her father and paternal grandmother had high serum aminopeptidase N (CD13; EC 3.4.11.2) activity (autosomal dominant). The molecular mass of the serum CD13 polypeptide of the propositus was larger than that of normal CD13, suggesting either a m...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:2001-02-01 00:00:00
abstract::Total protein (TP), albumin (Alb), transferrin (TRF), retinol-binding protein (RPB), N-acetyl-beta-glucosaminidase (NAG), alanine aminopeptidase (AAP), gamma-glutamyltransferase (GGT), and creatinine (Cr) were measured in random (untimed) urine samples from 29 nonpregnant women and from pregnant subjects (11 in the fi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-09-01 00:00:00
abstract:BACKGROUND:Detection of circulating tumor cells (CTCs) in the peripheral blood is a rapidly developing research field with clear clinical implications for the staging and monitoring of cancer patients. Current CTC assays, including the US Food and Drug Administration-cleared CellSearch® system, typically use markers [e...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2011.175570
更新日期:2012-05-01 00:00:00
abstract::We assessed several methods of inorganic phosphate assay for their suitability in estimating phospholipids in digested extracts of amniotic fluids. The extraction and digestion procedures used for phospholipids from amniotic fluid were also examined critically. The effect of contamination by blood or obstetric cream h...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-09-01 00:00:00
abstract::We describe a liquid-chromatographic assay for AZT in human plasma and urine. This assay involves the use of two internal standards, allowing reference of AZT peaks to the appropriate internal standard, the choice depending on the range of concentrations encountered. This method is isocratic, specific, sensitive enoug...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-08-01 00:00:00
abstract::Radioimmunoassays for haloperidol are described, involving use of tritium- or 125I-labeled drug or tritium-labeled spiroperidol, and a rabbit antiserum to a drug/bovine serum albumin conjugate. The 125I-labeled drug was prepared by the Chloramine T iodination technique. A fluoroimmunoassay for haloperidol is also desc...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1981-07-01 00:00:00
abstract::We find lag phases exceeding 20 min in measuring creatine kinase activity, by using the kinetic creatin phosphate leads to creatine assay, in sera of some patients with carcinoma metastatic to the liver. Such long lag phases are accompanied by a decreased apparent enzyme activity. These problems are eliminated by addi...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-11-01 00:00:00
abstract::A continuous-monitoring method for measuring acid phosphatase activity with alpha-naphthyl phosphate as the substrate was critically evaluated and modified. Using partially purified prostatic acid phosphatase, we show that certain conditions for the assay must be satisfied to ensure linearity. These conditions include...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-12-01 00:00:00
abstract:BACKGROUND:High-sensitivity C-reactive protein (hsCRP) testing is relatively inexpensive and has been shown to predict the risk of cardiovascular disease (CVD) and diabetes in multiple patient groups, including those treated with statin therapy. JUPITER (Justification for the Use of Statins in Primary Prevention: An In...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2008.109728
更新日期:2009-02-01 00:00:00
abstract:BACKGROUND:Kallikrein-related peptidase 6 (KLK6), a member of the serine protease family of kallikrein (KLK) genes, is dysregulated in ovarian carcinomas (OCa) and its overexpression is associated with poor prognosis. Regulation of its expression is poorly understood and is likely to be influenced by multiple mechanism...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2012.193060
更新日期:2013-01-01 00:00:00
abstract::We describe a simple polarization fluoroimmunoassay for the detection of several commonly abused drugs in urine. The single reagent used, prepared by mixing four different antisera and three fluorescein-labeled derivatives, produces a polarization signal that is the average of the individual signals of the derivatives...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00
abstract::Investigating the interference of danazol in testosterone radioimmunoassays, we measured serum testosterone concentrations in the serum of women receiving danazol, with three testosterone radioimmunoassay kits. All of these showed positive interference by danazol or its metabolites (or both). Values for testosterone o...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1983-01-01 00:00:00
abstract::We describe a time-resolved immunofluorometric assay (IFMA) for corticotropin in unextracted human plasma, based on the use of two monoclonal antibodies: europium-labeled antibody 1A12 and antibody 2A3 coated onto microtiter wells. We compared the results of this assay with those of an immunoradiometric assay (IRMA) p...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00
abstract::We report here the evaluation of the Abbott TDx assay with a monoclonal antibody for selectively quantifying cyclosporine (CsA) in whole blood. Over the clinically relevant concentration ranges, results with this assay demonstrated within- and between-run CVs of less than 2.5% and 5%, respectively; sensitivity of 25 m...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-11-01 00:00:00
abstract:BACKGROUND:Meconium analysis is a diagnostically sensitive and objective alternative to maternal self-report for detecting prenatal tobacco exposure. Nicotine and metabolite disposition in meconium is poorly characterized, and correlation of analytes' concentrations with neonatal outcomes is unexplored. Our objectives ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2008.109173
更新日期:2008-12-01 00:00:00
abstract::In this method for serum and urinary urea determination, the same reagent is used without predilution of urine samples. The method is based on the pH increase resulting from the ammonia released by urease hydrolysis of urea. o-Cresolphthalein complexone is used to monitor the pH change colorimetrically. Urea concentra...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-05-01 00:00:00
abstract::To validate azotemic markers as an index for intradialytic changes in solute concentration, we compared eight solutes (pseudouridine, xanthine, hypoxanthine, peak 4, peak 5, p-hydroxyhippuric acid, indoxyl sulfate, and hippuric acid) with five classical azotemic markers (urea, creatinine, uric acid, phosphate, and pot...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1992-08-01 00:00:00
abstract::We describe a modified tantalum ribbon atomic absorption procedure for determining lead in undiluted whole blood. An instrumentation Laboratory (I.L.) Model 151 atomic absorption spectrophotometer equipped with an I.L. Model 355 Flameless Sampler was used. The Flameless Sampler was slightly modified to include three-c...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-07-01 00:00:00
abstract::We examined the variability of lipid and lipoprotein concentrations in plasma from a population sample from the Jerusalem Lipid Research Clinic study. Coefficients of variation of about 8% for plasma cholesterol, 11% to 15% for low- and high-density-lipoprotein cholesterol, and about 30% for triglyceride were reported...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1985-07-01 00:00:00
abstract::We describe an enzyme-linked two-site immunoassay for quantitation of intact insulin in human serum and plasma. The method uses two murine monoclonal antibodies that bind to two different epitopes on the insulin molecule. The immunoassay is specific. Human proinsulin is not bound by the antibodies, and the binding of ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1993-04-01 00:00:00
abstract::We describe a rapid, sensitive, and specific "high performance" liquid chromatographic analysis for disopyramide and its mono-N-dealkylated metabolite in serum, urine, and saliva. We used a mu-Bondapak CN column and an acetate buffer mobile phase containing methanol. Retention times for the two compounds and the inter...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-03-01 00:00:00
abstract::The fructosamine test is considered clinically useful for assessing short-term integrated control of blood glucose, but there are few published data to support this hypothesis. We fractionated glycated and nonglycated proteins by affinity chromatography on phenylboronate columns and, with specific immunochemical metho...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-10-01 00:00:00