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

Éva Katona; Gizella Haramura; Levente Kárpáti; József Fachet; László Muszbek

doi:10.1055/s-0037-1613798

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2000; 83(02): 268-273
DOI: 10.1055/s-0037-1613798

Rapid Communication

Schattauer GmbH

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

Éva Katona

¹From the Department of Pathophysiology, University Medical School of Debrecen, Hungary

,

Gizella Haramura

²Department of Clinical Biochemistry and Molecular Pathology, University Medical School of Debrecen, Hungary

,

Levente Kárpáti

²Department of Clinical Biochemistry and Molecular Pathology, University Medical School of Debrecen, Hungary

,

József Fachet

¹From the Department of Pathophysiology, University Medical School of Debrecen, Hungary

,

László Muszbek

²Department of Clinical Biochemistry and Molecular Pathology, University Medical School of Debrecen, Hungary

› Author Affiliations

Abstract

Summary

A 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.

Keywords

Factor XIII - factor XIII ELISA - factor XIII reference interval

Full Text

References

References
1 Muszbek L, Ádány R, Mikkola H. Novel aspects of blood coagulation factor XIII. I. Structure, distribution, activation, and function. Crit Rev Clin Lab Sci 1996; 33: 357-421.
2 Saito M, Asakura H, Yoshida T, Ito K, Okafuji K, Yoshida T, Matsuda T. A familial factor XIII subunit B deficiency. Br J Haematol 1990; 74: 290-4.
3 Tosetto A, Castaman G, Rodeghiero F. Abnormal plasma factor XIII subunit A with fully functional platelet FXIII in a family with congenital deficiency of FXIII subunit B. Thromb Haemost 1993; 69: 1295a.
4 Izumi T, Hashiguchi T, Castaman G, Tosetto A, Rodeghiero F, Girolami A, Ichinose A. Type I factor XIII deficiency is caused by a genetic defect of its B subunit: Insertion of triplet AAC in exon III leads to premature termination in the second Sushi domain. Blood 1996; 87: 2769-74.
5 Lorand L, Losowsky MS, Miloszewski KJA. Human factor XIII: fibrin-stabilising factor. Prog Haemost Thromb 1980; 05: 245-90.
6 Board PG, Losowsky MS, Miloszewski KJA. Factor XIII: inherited and acquired deficiency. Blood Rev 1993; 07: 229-42.
7 Egbring R, Kröniger A. Seitz Factor XIII deficiency: Pathogenic mechanisms and clinical significance. Sem Thromb Hemost 1996; 22: 419-25.
8 Muszbek L, Polgár J, Fésüs L. Kinetic determination of blood coagulation factor XIII in plasma. Clin Chem 1985; 31: 35-40.
9 Fickenscher K, Aab A, Stuber W. A photometric assay for blood coagulation factor XIII. Thromb Haemost 1991; 65: 535-40.
10 Lorand L, Urayama T, deKiewiet JWC, Nossel HL. Diagnostic and genetic studies on fibrin-stabilising factor with a new assay based on amine incorporation. J Clin Invest 1969; 48: 1054-64.
11 Lorand L, Campbell-Wilkes LK, Cooperstein L. A filter paper assay for transamidating enzymes using radioactive amine substrates. Anal Biochem 1972; 50: 623-31.
12 Song YC, Sheng D, Taubenfeld SM, Matsueda GR. A microtiter assay for factor XIII using fibrinogen and biotinylcadaverine as substrates. Anal Biochem 1994; 223: 88-92.
13 Barbui T, Rodeghiero F, Dini E, Mariani G, Papa ML, de Biasi R, Cordero RMurillo, Montero CUmana. Subunit A and S inheritance in four families with congenital factor XIII deficiency. Brit J Haematol 1978; 38: 267-71.
14 Skrzynia C, Reisner HM, McDonagh J. Characterisation of the catalytic subunit of factor XIII by radioimmunoassay. Blood 1982; 60: 1089-95.
15 Ikematsu S, McDonagh RP, Reisner HM, Skrzynia C, McDonagh J. Immunochemical studies of human factor XIII. Radioimmunoassay for the carrier subunit of the zymogen. J Lab Clin Med 1981; 97: 662-71.
16 Yorifuji H, Andersson K, Lynch GV, Van De Water L, McDonagh J. B protein of factor XIII: Differentiation between free B and complexed B. Blood 1988; 72: 1645-50.
17 Murdock PJ, Owens DL, Chitolie A, Hutton RA, Lee CA. Development and evaluation of ELISAs for factor XIIIA and XIIIB subunits in plasma. Thromb Res 1992; 67: 73-9.
18 Lorand L, Credo RB, Janus TJ. Factor XIII (fibrin stabilizing factor). Methods Enzymol 1981; 80: 333-41.
19 Schwartz ML, Pizzo SV, Hill RL, McKee PA. Human factor XIII from plasma and platelets. J Biol Chem 1973; 248: 1395-407.
20 McDonagh J, Waggoner WG, Hamilton EG, Hindenach B, McDonagh RP. Affinity chromatography of human plasma and platelet factor XIII on organomercurial agarose. Biochim Biophys Acta 1976; 446: 345-57.
21 Bohn H, Schwick HG. Isolierung und Charakterisierung eines fibrinstabilisierenden Faktors aus menschlichen Plazenten. Arzneim Forsch 1971; 21: 1432-9.
22 Polgár J, Hidasi V, Muszbek L. Non-proteolytic activation of cellular protransglutaminase (placental macrophage factor XIII). Biochem J 1990; 267: 557-60.
23 Stahli C, Staehelin T, Miggiano V, Schmidt J, Haring P. High frequencies of antigen specific hybridomas: Dependence on immunisation parameters and prediction by spleen cell analysis. J Immunol Methods 1980; 32: 297-309.
24 Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody predefined specificity. Nature 1975; 256: 495-506.
25 O’Shannessy DJ. Antibodies biotinylated via sugar moieties. Methods Enzymol 1990; 184: 162-6.
26 Wilson MB, Nakane PK. Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: Immunofluorescence Related Staining Techniques. Knapp W, Holubar K, Wick G. Amsterdam: Elsevier; 1978: 215-20.
27 Kárpáti L, Muszbek L. An improved photometric assay for the determination of blood coagulation factor XIII in plasma. International Congress on Pediatric Laboratory Medicine Abstract Book 1998; 29.
28 Kohler HP, Stickland Ossei-Gerning N, Carter A, Mikkola H, Grant P. Association of a common polymorphism in the factor XIII gene with myocardial infarction. Thromb Haemost 1998; 79: 8-13.
29 Catto JA, Kohler HP, Coore J, Mansfield MW, Stickland MH, Grant P. Association of a common polymorphism in the factor XIII gene with venous thrombosis. Blood 1999; 93: 906-8.
30 Seitz R, Duckert S, Lopaciuk S, Muszbek L, Rodeghiero F, Seligsohn U. Study Group ETRO Working Party on factor XIII questionnaire on congenital factor XIII deficiency in Europe: status and perspectives. Sem Thromb Hemost 1996; 22: 415-8.
31 Anwar R, Gallivan L, Edmonds SD, Markham AF. Genotype/phenotype correlations for coagulation factor XIII: specific normal polymorphisms are associated with high or low factor XIII specific activity. Blood 1999; 93: 897-905.
32 Greenberg CS, Shuman MA. The zymogen form of blood coagulation factor XIII binds specifically to fibrinogen. J Biol Chem 1982; 257: 6096-101.
33 Mikkola H, Muszbek L, Laiho E, Syrjälä M, Hämäläinen E, Haramura G, Salmi T, Peltonen L, Palotie A. Molecular mechanism of a mild phenotype in coagulation factor XIII (FXIII) deficiency: a splicing mutation permitting partial correct splicing of FXIII A-subunit mRNA. Blood 1997; 89: 1279-87.
34 Miloszewski KJA, Losowsky MS. The half life of factor XIII in vivo. Br J Haematol 1978; 38: 267-71.
35 Fear JD, Miloszewski KJA, Losowsky MS. The half life of factor XIII in the management of inherited deficiency. Thromb Haemost 1983; 49: 102-5.
36 Rodeghiero F, Tosetto A, Di Bona E, Castaman G. Clinical pharmacokinetics of placenta derived factor XIII concentrate in type I and type II factor XIII deficiency. Am J Haematol 1991; 36: 30-4.