A Photometric Assay for Blood Coagulation Factor XIII

1991 ◽  
Vol 65 (05) ◽  
pp. 535-540 ◽  
Author(s):  
Karl Fickenscher ◽  
Angela Aab ◽  
Werner Stüber
Keyword(s):  
Factor Xiii ◽  
Coagulation Factor ◽  
Photometric Determination ◽  
Peptide Substrate ◽  
Normal Range ◽  
Step Procedure ◽  
Healthy Donors ◽  
One Step ◽  
Aggregation Inhibitor

SummaryAn assay for a direct photometric determination of F XIII in untreated and undiluted plasma was developed. In a one-step procedure F XIII is activated by thrombin and Ca2+ and crosslinks glycine-ethylester to a specific glutamine containing peptide substrate. The released ammonia is incorporated into α-keto-glutarate by glutamate dehydrogenase, and the NADH consumption of this reaction is measured photometrically at 340 nm. NADH-consumption is directly proportional to the F XIII activity. Fibrin polymerization and the corresponding turbidity is avoided by the use of a fibrin aggregation inhibitor.The procedure is rapid and simple and enables to measure within the range of 0 to 150% F XIII. It can be performed with automated analyzers as well as with common photometric equipment.The normal range of F XIII activity in 167 healthy donors was determined to be 70 to 140%.

Improved rapid assay of uric acid in serum by liquid chromatography.

1988 ◽  
Vol 34 (12) ◽  
pp. 2567-2568 ◽  
Author(s):  
M Tanaka ◽  
M Hama
Keyword(s):  
Uric Acid ◽  
High Performance ◽  
Rapid Determination ◽  
Correlation Coefficients ◽  
Gel Permeation Chromatography ◽  
Serum Samples ◽  
Step Procedure ◽  
One Step ◽  
Highly Porous

Abstract This improved method for rapid determination of uric acid in serum is based on high-performance liquid-gel-permeation chromatography, with hydrophilic and highly porous vinyl alcohol copolymer as packing material. It has the following advantages: no need for sample deproteinization or use of a precolumn, more than 500 serum samples can be analyzed without having to regenerate or recondition the analytical apparatus, and the analysis for uric acid is a one-step procedure. Correlation coefficients between this method and other methods are very good (r = 0.998, 0.999).

The Val34Leu Polymorphism in the A Subunit of Coagulation Factor XIII Contributes to the Large Normal Range in Activity and Demonstrates That the Activation Peptide Plays a Role in Catalytic Activity

Blood ◽  
1998 ◽  
Vol 92 (8) ◽  
pp. 2766-2770 ◽  
Author(s):  
S. Kangsadalampai ◽  
P.G. Board
Keyword(s):  
Factor Xiii ◽  
Direct Evidence ◽  
Coagulation Factor ◽  
Normal Range ◽  
Thrombin Cleavage ◽  
Coagulation Factor Xiii ◽  
A Subunit ◽  
Elevated Activity ◽  
Activation Peptide ◽  
American Society

There is a wide normal range of coagulation factor XIII activity that has never been adequately explained. A polymorphism substituting leucine for valine at position 34 in the activation peptide of the A subunit of factor XIII has recently been discovered in nondeficient individuals, and the present studies indicate that the leucine substitution results in a significant increase in transglutaminase activity. The frequency of the Leu34 allele in the Australian Caucasian population is 0.27, which is high enough to suggest that the inheritance of either the Val34 or Leu34 alleles may contribute to the wide normal range of activity. Although there has been structural evidence indicating that the activation peptide does not dissociate from the enzyme after thrombin cleavage, the discovery of elevated activity resulting from the Leu34 substitution is the first direct evidence that the activation peptide plays a continuing role in the function of factor XIII. © 1998 by The American Society of Hematology.

Functional analysis of fibrin γ-chain cross-linking by activated factor XIII: determination of a cross-linking pattern that maximizes clot stiffness

Blood ◽  
2007 ◽  
Vol 110 (3) ◽  
pp. 902-907 ◽  
Author(s):  
Kristina F. Standeven ◽  
Angela M. Carter ◽  
Peter J. Grant ◽  
John W. Weisel ◽  
Irina Chernysh ◽  
...  
Keyword(s):  
Factor Xiii ◽  
Tissue Transglutaminase ◽  
Coagulation Factor ◽  
Fold Increase ◽  
Cross Linking ◽  
Cross Links ◽  
Α Chain ◽  
And Function ◽  
Hybrid Cross

Abstract Activated coagulation factor XIII (FXIIIa) cross-links the γ-chains of fibrin early in clot formation. Cross-linking of the α-chains occurs more slowly, leading to high molecular weight multimer formations that can also contain γ-chains. To study the contribution of FXIIIa-induced γ-chain cross-linking on fibrin structure and function, we created 2 recombinant fibrinogens (γQ398N/Q399N/K406R and γK406R) that modify the γ-chain cross-linking process. In γK406R, γ-dimer cross-links were absent, but FXIIIa produced a cross-linking pattern similar to that observed in tissue transglutaminase cross-linked fibrin(ogen) with mainly α-γ cross-links. In Q398N/Q399N/K406R, cross-links with any γ-chain involvement were completely absent, and only α-chain cross-linking occurred. Upon cross-linking, recombinant normal fibrin yielded a 3.5-fold increase in stiffness, compared with a 2.5-fold increase by α-chain cross-linking alone (γQ398N/Q399N/K406R). γK406R fibrin showed a 1.5-fold increase in stiffness after cross-linking. No major differences in clot morphology, polymerization, and lysis rates were observed, although fiber diameter was slightly lower in cross-linked normal fibrin relative to the variants. Our results show that γ-chain cross-linking contributes significantly to clot stiffness, in particular through γ-dimer formation; α-γ hybrid cross-links had the smallest impact on clot stiffness.

A highly sensitive fluorometric assay for determination of human coagulation factor XIII in plasma

2007 ◽  
Vol 367 (2) ◽  
pp. 152-158 ◽  
Author(s):  
Kai Oertel ◽  
Andreas Hunfeld ◽  
Edgar Specker ◽  
Christine Reiff ◽  
Rainer Seitz ◽  
...  
Keyword(s):  
Factor Xiii ◽  
Coagulation Factor ◽  
Fluorometric Assay ◽  
Coagulation Factor Xiii ◽  
Highly Sensitive ◽  
Human Coagulation Factor Xiii

A Simple, Quick One-step ELISA Assay for the Determination of Complex Plasma Factor XIII (A2B2)

2000 ◽  
Vol 83 (02) ◽  
pp. 268-273 ◽  
Author(s):  
Éva Katona ◽  
Gizella Haramura ◽  
Levente Kárpáti ◽  
József Fachet ◽  
László Muszbek
Keyword(s):  
Factor Xiii ◽  
Sandwich Elisa ◽  
High Sensitivity ◽  
Reference Interval ◽  
Elisa Assay ◽  
Plasma Factor ◽  
Sandwich Elisa Assay ◽  
One Step ◽  
A Subunit

SummaryA new highly sensitive sandwich ELISA assay was developed for the determination of plasma factor XIII (FXIII). Plasma FXIII is a tetrameric complex of two types of subunits (A2B2). A biotinylated monoclonal capture-antibody against the B-subunit and a peroxidase-labelled monoclonal tag-antibody against the A-subunit were added to the plasma dilution and the amount of the complex attached to streptavidincoated microplate was quantitated by measuring peroxidase activity. Only the tetrameric plasma FXIII reacted in the assay, non-complexed A or B subunits showed no reaction. The assay is linear up-to 40 µg/L of FXIII in the assay mixture. It is a quick one-step assay which can be performed within two hours. At normal and low FXIII concentration within batch reproducibility was 2.0% and 3.3%, day to day variation was 5.5% and 8.7%, respectively. Its high sensitivity allows reliable measurement at FXIII concentrations below 1% of normal average. Plasma samples can be stored for the assay at –20° C for at least one month. Plasma levels of healthy individuals were normally distributed and no gender difference was observed. A reference interval of 14-28 mg/L (67-133%) was established.

Relation between ADAMTS13 activity and ADAMTS13 antigen levels in healthy donors and patients with thrombotic microangiopathies (TMA)

2006 ◽  
Vol 95 (02) ◽  
pp. 212-220 ◽  
Author(s):  
Johanna A. Hovinga ◽  
Christian Konetschny ◽  
Andrea Herzog ◽  
Letizia Koller ◽  
Alfred Weber ◽  
...  
Keyword(s):  
Rapid Determination ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Adamts13 Activity ◽  
Normal Range ◽  
Thrombotic Microangiopathies ◽  
Present Evidence ◽  
Antigen Elisa ◽  
Healthy Donors

SummaryWe have established a new, enzyme-linked immunosorbent assay (ELISA) for the detection of ADAMTS13 antigen using purified polyclonal rabbit anti-human ADAMTS13 IgG. Normal plasma ADAMTS13 antigen levels span a concentration of 740 –1420ng/ml (median 1080ng/ml) resulting in an ADAMTS13 activity to antigen ratio of 0.48 to 1.68 U/µg. In a cohort of HUS patients, ADAMTS13 antigen was in the normal range, whereas in hereditary TTP patients antigen levels were low to undetectable, in concordance with severe deficient ADAMTS13 activity. Plasma of acquired TTP patients was found to contain free as well as autoantibody-bound ADAMTS13. We also present evidence for circulating anti-ADAMTS13 antibody/ADAMTS13 antigen immune complexes not only in acutely ill or actively treated patients but also in patients who have already achieved clinical remission. This new developed ADAMTS13 antigen ELISA assay allows rapid determination of ADAMTS13 antigen levels in human plasma but is of limited predictive value for the diagnosis or treatment of acquired TTP due to the detection of ADAMTS13 in antibody complexes.

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