Fibronectin quantification without antibodies

A bioassay for the detection of the gelatin-captured macromolecule

Ágnes Pataki, E. Madarász, István Kurucz

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

We have developed a new cell-adhesion-bioassay (CAA) for the quantitative determination of fibronectin in biological fluids. The assay is based on two particular properties of fibronectin: it specifically binds to gelatin with high affinity and simultaneously it can anchor to different surface molecules of a cell. First fibronectin, derived from very different biological fluids, is purified in situ, within the wells of the microtiter plates applied for the assay, using solid surface bound gelatin. After capturing the macromolecule, it is quantified based on its cell adhesive properties. In contrast to ELISA the CAA does not require specific antibodies, and as the Jurkat cells used as indicator cells, seem to recognize fibronectin from different species equally; species specificity of the reagent plays smaller, perhaps negligible, role in the determination of the amount of the macromolecule. The CAA method may not replace fibronectin specific ELISA-s, but using its principle, improved applications, for example a capture EIA for determining fibronectin can easily be envisioned and CAA may serve as a viable alternative for EIA-s when specific antibodies are not available or when relative measurement of not only the soluble but cell surface associated fibronectin is necessary.

Original languageEnglish
Pages (from-to)113-126
Number of pages14
JournalJournal of Biochemical and Biophysical Methods
Volume68
Issue number2
DOIs
Publication statusPublished - Aug 31 2006

Fingerprint

Bioassay
Gelatin
Macromolecules
Fibronectins
Biological Assay
Cell adhesion
Antibodies
Cell Adhesion
Assays
Enzyme-Linked Immunosorbent Assay
Species Specificity
Jurkat Cells
Fluids
Anchors
Adhesives
Molecules

Keywords

  • CAA
  • Cell-adhesion
  • ELISA
  • Fibronectin-assay
  • Gelatin-captured
  • Integrin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics

Cite this

Fibronectin quantification without antibodies : A bioassay for the detection of the gelatin-captured macromolecule. / Pataki, Ágnes; Madarász, E.; Kurucz, István.

In: Journal of Biochemical and Biophysical Methods, Vol. 68, No. 2, 31.08.2006, p. 113-126.

Research output: Contribution to journalArticle

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