Abstract:
:Solid-phase-based immunoassays have traditionally been plagued by nonspecific binding to the solid phase and by slow reaction kinetics relative to reactants that are free to diffuse in solution. We have developed two novel immunoassays in which the solid phase is generated in situ after the specific binding reaction has occurred, thereby enhancing reaction kinetics and minimizing the opportunities for non-specific binding. In the first system, the capture antibody is conjugated to an organic monomer, polymerization of which to form insoluble polymer particles is initiated by a reaction involving free radicals. The amount of signal-labeled antibody incorporated into the resulting particles is directly proportional to the concentration of antigen. The principle is illustrated for the simultaneous assay of IgG and IgM in a single sample. In the second system, capture antibody is conjugated to a polymer, the solubility of which is a function of temperature. Specific binding is conducted below the critical solution temperature of the polymer, which is then separated from solution by increasing the temperature above the critical temperature. The incorporation of signal-labeled antibody into the precipitated polymer is directly proportional to the concentration of antigen. This principle is illustrated for the assay of hepatitis B surface antigen and Chlamydia trachomatis.
journal_name
Clin Chemjournal_title
Clinical chemistryauthors
Auditore-Hargreaves K,Houghton RL,Monji N,Priest JH,Hoffman AS,Nowinski RCsubject
Has Abstractpub_date
1987-09-01 00:00:00pages
1509-16issue
9eissn
0009-9147issn
1530-8561journal_volume
33pub_type
杂志文章abstract::To understand the regulation of immunity one must understand the processes and the consequences of the activation of the lymphoid cell, its clonal proliferation, and the ultimate differentiation of the progeny into the effector elements that constitute the immune system. By use of polyclonal mitogens, individual T-cel...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-12-01 00:00:00
abstract::Diabetic patients in poor glycemic control show increased glycation of total plasma proteins, but little is yet known about the relative extents to which the various individual proteins are glycated. Thus, we studied the non-enzymic glycation of several major plasma proteins and plasma protein fractions in normal and ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-12-01 00:00:00
abstract::The manual Reference Method of the Centers for Disease Control for serum iron (CDC/RM/Fe) and a semiautomated adaptation of it were used to evaluate two working methods: one, a detergent solubilization procedure for the Roche Cobas-Bio analyzer, the other, the Kodak Ektachem 700 procedure, based on dry-film technology...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-10-01 00:00:00
abstract::We report a simple, enzymatic, end point method for determining nitrate in serum and urine with use of nitrate reductase from Aspergillus sp. (EC 1.6.6.2). The decrease in absorbance at 340 nm as a result of the oxidation of beta-NADPH is measured. We used FAD as a supplementary electron carrier and added an internal ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1995-06-01 00:00:00
abstract::We investigated two techniques of immunoblotting--the Western blot and the dot blot--for use in detecting prostatic acid phosphatase (PAP, EC 3.1.3.2). We used polyclonal antisera to human PAP, produced in rabbits by hyperimmunization with purified PAP, and PAP-specific monoclonal antibodies in the immunoenzymatic pro...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
abstract::One currently observes many healthcare institutions rushing to reengineer and install information systems with the expectation of achieving enhanced efficiency, competitiveness, and, it is hoped, higher patient satisfaction resulting from timely, high-quality care. Unfortunately, information system concepts, design, a...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1995-09-01 00:00:00
abstract:BACKGROUND:Recent studies show that the haptoglobin phenotype in individuals with diabetes mellitus is an important factor for predicting the risk of myocardial infarction, cardiovascular death, and stroke. Current methods for haptoglobin phenotyping include PCR and gel electrophoresis. A need exists for a reliable met...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2015.244004
更新日期:2016-01-01 00:00:00
abstract::Phenobarbital and phenytoin are extracted and concentrated in a single step, without solvent evaporation, followed by on-column methylation and gas-chromatographic quanitation. A similar extraction step for the assay of theophylline by high-pressure liquid chromatography is described. The extraction step can be comple...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1978-09-01 00:00:00
abstract::We describe a chromatographic procedure for sensitive (2.0 mumol/L), specific quantification of vanillylmandelic acid (VMA) in urine with iso-VMA as an internal standard. After rapid extraction from urine, the VMA is determined by isocratic reversed-phase ion-pair chromatography on a bonded C8 column and detection of ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-04-01 00:00:00
abstract::Organic anions accumulated in blood serum of patients with chronic renal failure were separated by a novel technique: closed-system capillary zone electrophoresis (CZE) in a pH6 carrier-electrolyte system. Hippuric acid (HA), p-hydroxyhippuric acid, and uric acid were identified by their co-elution with standards prep...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1990-03-01 00:00:00
abstract::This highly sensitive method for determining retinol-binding protein in human serum and urine is based on a double-antibody "sandwich"-type enzyme-linked immunosorbent assay. The assayable concentration range is 0.8-48 micrograms/L, the detection limit 0.2 micrograms/L. Within-assay coefficients of variation for 10 de...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1984-01-01 00:00:00
abstract::We describe a new multilayer immunoassay element for the determination of haptens in undiluted serum and plasma. Polysaccharide layers are coated onto a plastic base. The signal layer contains an immobilized antibody and a fluorescent-labeled hapten. A second layer, containing a pigment, acts as an optical screen. Sam...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-09-01 00:00:00
abstract::We describe a method used for determining serotonin and tryptamine in urine after oral loading with their precursor amino acid. Several factors affecting prepurification and chromatographic separation of both indoleamines were optimized, including sample extraction and mobile-phase composition. Under optimal condition...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1989-01-01 00:00:00
abstract::Analytical, intra-individual, and interindividual components of variation have been determined for pH, pCO2, bicarbonate, base excess, and total CO2 in capillary specimens of whole blood from a cohort of 14 healthy subjects. Calculated analytical goals (SD) are pH less than or equal to 0.008, pCO2 less than or equal t...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1987-08-01 00:00:00
abstract:BACKGROUND:The kidney performs a multitude of essential functions to maintain homeostasis. In clinical medicine, glomerular filtration rate (GFR) provides the best index of overall kidney function, and proteinuria adds additional information on renal and nonrenal prognosis. Several novel biomarkers of kidney injury and...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2011.167494
更新日期:2012-04-01 00:00:00
abstract::We describe a convenient, economical procedure for measuring uroporphyrinogen I synthase (EC 4.3 1.8) activity in erythrocytes, the results of which can be used to diagnose acute intermittent porphyria, in either its latent or acute stage. By limiting the test reaction sequence to the conversion of porphobilinogen to ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1980-07-01 00:00:00
abstract::We used ion-exchange column chromatography and electrophoresis on polyacrylamide gel to compare the acid phosphatase isoenzymes of prostate and leukocytes. The major isoenzyme of the prostate is band 2A; only a trace of band 2B was observed. However, the major isoenzymes of leukocytes are band 4 and band 2B, and only ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1979-07-01 00:00:00
abstract::Energy-dispersive x-ray spectroscopy and inductively coupled plasma emission spectrometry were evaluated as methods for routine multielement analysis of biological material. Standard samples included Standard Reference Materials (National Bureau of Standards), compounded mixtures, and supplements that provided a wide ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1976-12-01 00:00:00
abstract:BACKGROUND:Recent publications have explored the possibility of using fingerprints to confirm drug use, but none has yet dealt with environmental contamination from fingertips. Here we explored the possibility of establishing an environmental cutoff for drug testing from a single fingerprint. METHODS:Fingerprint sampl...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2017.281469
更新日期:2018-06-01 00:00:00
abstract:BACKGROUND:Simple, reproducible assays are needed for the quantification of sphingolipids, ceramide (Cer), and sphingoid bases. We developed an HPLC method for simultaneous quantification of total plasma concentrations of Cer, glucosylceramide (GlcCer), and ceramide trihexoside (CTH). METHODS:After addition of sphinga...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2006.079012
更新日期:2007-04-01 00:00:00
abstract:BACKGROUND:First introduced in 2006, recombinase polymerase amplification (RPA) has stirred great interest, as evidenced by 75 publications as of October 2015, with 56 of them just in the last 2 years. The widespread adoption of this isothermal molecular tool in many diagnostic fields represents an affordable (approxim...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:10.1373/clinchem.2015.245829
更新日期:2016-07-01 00:00:00
abstract::To determine the molar absorptivities of beta-NADH and beta-NAD at 260 nm, we purified the reduced form of the coenzyme by repeated anion-exchange chromatography. A NADH preparation was so obtained for which the 260 nm/340 nm absorbance ratio was 2.265. When calculated with epsilon 340 beta-NADH = 6.22 times 10-3 for ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-06-01 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:BACKGROUND:Newborn screening for deficiency in the lysosomal enzymes that cause Fabry, Gaucher, Krabbe, Niemann-Pick A/B, and Pompe diseases is warranted because treatment for these syndromes is now available or anticipated in the near feature. We describe a multiplex screening method for all five lysosomal enzymes tha...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:10.1373/clinchem.2004.035907
更新日期:2004-10-01 00:00:00
abstract:BACKGROUND:Studies of the effects of tobacco smoke often rely on reported exposure to cigarette smoke, a measure that is subject to bias. We describe here the relationship between parental smoking exposure as assessed by urinary cotinine excretion and lung function in children with asthma. METHODS:We studied 90 childr...
journal_title:Clinical chemistry
pub_type: 临床试验,杂志文章
doi:
更新日期:1999-04-01 00:00:00
abstract::Physiologic systems have substantial reserves in younger individuals. The process of aging and intercurrent pathologic processes gradually eliminate these reserves. Changes in endocrine systems, including menopause in women, androgen deficiency in men, loss of skeletal mass, decrease in growth hormone serum concentrat...
journal_title:Clinical chemistry
pub_type: 杂志文章,评审
doi:
更新日期:1999-08-01 00:00:00
abstract::Detection and characterization of small IgM paraproteins by immunoelectrophoresis are often difficult because of (a) slow diffusion of the macromolecular immunoglobulin, and (b) the obscuring of the light chain, especially kappa, by the light chains of background polyclonal immunoglobulin (the "umbrella effect"). Vari...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1988-08-01 00:00:00
abstract::In this new radioimmunoassay system for determination of amatoxins in urine and plasma, a novel chemical approach is used for antigen and 125I-tracer production, based on a detoxified alpha-amanitin derivative (aldoamanitin). Total assay time, including data processing, is less than 100 min. The lowest detectable conc...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1986-09-01 00:00:00
abstract:BACKGROUND:Prediction of cyclosporine (CSA) efficacy and toxicity in individual patients is difficult. There is no practical, biologically relevant, pharmacodynamic measure of CSA effect. A major effect of CSA is to decrease interleukin-2 (IL-2) production; however, measurement of this effect in isolated lymphocytes as...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1999-09-01 00:00:00
abstract::With use of a specially designed gradient-generating system, of anion-exchange resins of 10-20 mum particle diameter, and of an orcinol solution in concentrated sulfuric acid (1 g/liter) as detection reagent, a mixture of 16 carbohydrates can be separated in less than 4 h. The limit of detection for most sugars is in ...
journal_title:Clinical chemistry
pub_type: 杂志文章
doi:
更新日期:1975-12-01 00:00:00