Automated Two-Stage Assay for Determination of Antithrombin III with a Centrifugal Analyzer

Abstract

The thrombin-inactivating α₂-globulin, antithrombin III (At-III), was measured in plasma and urine with a chromogenic tripeptide substrate, using a centrifugal analyzer. Plasma At-III was subnormal in liver disease. In acute myocardial infarction a mean At-III of approximately 120% of normal was found. At-III in persons who developed deep-vein thrombosis decreased significantly (p < 0.05) between the day of admission and the 3rd day after admission to hospital. Urine from normals did not contain detectable At-III. With increasing urinary albumin increasing concentration of At-III was found.

Automation of Antithrombin III Methods on Routinely Available Instruments

“…The choice of an instrument should be directed by the number of tests performed per day, the size of the workload (small or large daily series, large weekly series), as well as tests performed on the same instrument (coagulation or biochemistry). Choice of instrument also depends on whether tests will be performed routinely and 36 Dettori et al 20 Andreasen 6 Holmes et al 28 Brandt et al 11 Bartl and Lill 8 Scully and Kakkar 39 Conard et al 19 Cederholm-Williams 13 Holmes et al 28 Aiach et al 4 Droullé et al 21 Kahle et al 30 Bartl and Lill 8 Dettori et al 20 Bartl and Lill 8 Prellwitz et al 38 Frantzen et al 24 Fareed et al 23 Prellwitz et al 38 whether the biologist wants to be able to modify conditions of the tests. The improvement of the quality of the results and the rapidity of the instruments should allow large-scale surveys of AT III, such as for epidemiologic purposes.…”

“…In contrast, chromogenic methods are easily automated by kinetic rate technique, and most of the instruments used in clinical chemistry laboratories are appropriate. Results obtained with the following instruments have been published: GEMSAEC centrifugal analyzer, 6,20,36 Multistat microcentrifugal analyzer, 28 ABA-100 Abbott, 8,11 Vitatron Automated Kinetic Enzyme and Substrate Analyzer, 8,30 Gilford 3500, 13,19,28 Isamat Isabiologie, 4 Kem-O-Mat 2T Coultronics, 21 LKB, 39 Olli 3000, 20 Eppendorf 5020, 8 Cobas Bio, 24,38 and aca DuPont. 17,23,38 The ABA-100 is only semiautomated.…”

Antithrombin III and Related Parameters in Surgical Patients Receiving Blood Components

“…Therefore, we calculated and plotted one reference curve from 10 daily runs (standard plasma = 99%) and derived all functional activities from this curve. The normal average of the method which we used is stated lobe 100% and the range +2 SD 70-130% [4,27], Immunoreactive AT was determined by radial im munodiffusion with Behring M-Partigen® plates using citrate plasma and correcting for the dilution. The nor-mal average and range +2 SD given by the manufac turer are 30.5 and 22.0-30.0mg/dl, respectively, To facilitate comparisons with the functional assay, we transformed our results into percent of that normal average.…”

Contributions of Histochemistry to the Development of the Proteolytic Enzyme Detection System in Diagnostic Medicine

“…The above four assays do not require a high degree of sensitivity; consequently, activity in plasma can be determined colorimetrically (1,2) or with a moderately intense fluorophore (Figure 4). However, some proteases of the hemostatic system (11) and most tissue proteases (4) found in body fluids are present in relatively smaller quantities, even under pathological conditions, and require a much more sensitive detection system to be accurately quantitated (44).…”

“…It was Gomori, in 1954, who used chloroacetyl-/3-naphthylamine with ammonia to obtain glycine and D,L-alanyl-/3-naphthyl-'Presented as part of the program at the 1982 Joint Meeting of the American and Japanese Histochemical Societies, held in Vancouver, British Columbia, Canada, July [20][21][22][23][24]1982. 2 Portions of this work was performed under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory under contract number W-7405-ENG-48; other parts were carried out at Enzyme Systems Products Inc. as designated in the text.…”