A simple, quick one-step ELISA assay for the determination of complex plasma factor XII (A2B2)

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Abstract

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 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.

Original languageEnglish
Pages (from-to)268-273
Number of pages6
JournalThrombosis and Haemostasis
Volume83
Issue number2
Publication statusPublished - 2000

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Factor XII
Factor XIII
Enzyme-Linked Immunosorbent Assay
Peroxidase
Monoclonal Antibodies
Streptavidin

Keywords

  • Factor XIII
  • Factor XIII ELISA
  • Factor XIII reference interval

ASJC Scopus subject areas

  • Hematology

Cite this

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title = "A simple, quick one-step ELISA assay for the determination of complex plasma factor XII (A2B2)",
abstract = "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 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",
author = "E. Katona and G. Haramura and L. K{\'a}rp{\'a}ti and J{\'o}zsef Fachet and L. Muszbek",
year = "2000",
language = "English",
volume = "83",
pages = "268--273",
journal = "Thrombosis and Haemostasis",
issn = "0340-6245",
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T1 - A simple, quick one-step ELISA assay for the determination of complex plasma factor XII (A2B2)

AU - Katona, E.

AU - Haramura, G.

AU - Kárpáti, L.

AU - Fachet, József

AU - Muszbek, L.

PY - 2000

Y1 - 2000

N2 - 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 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.

AB - 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 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.

KW - Factor XIII

KW - Factor XIII ELISA

KW - Factor XIII reference interval

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