A simple, quick one-step ELISA assay for the determination of complex plasma factor XIII (A2B2).
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 streptavidin-coated 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 microg/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 degrees 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.
Katona
E
E
Department of Pathophysiology, University Medical School of Debrecen, Hungary.
Haramura
G
G
Kárpáti
L
L
Fachet
J
J
Muszbek
L
L
D000911:Antibodies, Monoclonal; D019921:Biotinylation; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards*; D005176:Factor XIII / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000493:pharmacokinetics; D005177:Factor XIII Deficiency / Q000097:blood / Q000188:drug therapy; D005340:Fibrinogen / Q000378:metabolism; D006735:Horseradish Peroxidase; D006801:Humans; D020836:Protein Structure, Quaternary; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D011503:Transglutaminases / Q000276:immunology
D000911:Antibodies, Monoclonal; D005340:Fibrinogen; D005176:Factor XIII; C054380:factor XIIIb; D006735:Horseradish Peroxidase; D011503:Transglutaminases