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Related Experiment Videos

Two sensitive time-resolved fluoroimmunoassays for cellular fibronectin

J Kropf1, A M Gressner

  • 1Abt. Klinische Chemie und Zentrallaboratorium, Philipps-Universität, Marburg, Germany.

Clinical Chemistry
|September 1, 1995
PubMed
Summary
This summary is machine-generated.

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Two new immunoassays accurately detect cellular fibronectin (cFN) in plasma and cell cultures. Assay 2 demonstrated superior parallelism and precision for reliable cFN quantification.

Area of Science:

  • Biochemistry
  • Immunology
  • Assay Development

Background:

  • Cellular fibronectin (cFN) is a crucial extracellular matrix protein.
  • Accurate quantification of cFN is vital for understanding cellular processes and disease states.

Purpose of the Study:

  • To develop and validate sensitive sandwich immunoassays for cellular fibronectin (cFN) detection.
  • To compare the performance of two distinct assay formats for cFN quantification in biological samples.

Main Methods:

  • Development of two sandwich immunoassays utilizing monoclonal antibodies specific to the EDA sequence of cFN.
  • Application of time-resolved fluorescence spectrophotometry for sensitive detection.
  • Testing in cell culture supernatants and human plasma samples.

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Main Results:

  • Both assays achieved detection limits between 2.6-4.0 µg/L for cFN.
  • Assay 2 exhibited excellent parallelism in human plasma over a wide concentration range (12-1000 µg/L).
  • Assay 2 showed better precision with lower between-run coefficients of variation (7.2%) compared to Assay 1 (11.4%).

Conclusions:

  • Sensitive and reliable immunoassays for cFN determination have been successfully developed.
  • Assay 2, with immobilized antibody, is recommended for accurate cFN quantification in human plasma due to its superior performance.
  • These assays provide valuable tools for research involving cellular fibronectin.