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Monoclonal antibodies with selective specificity towards different glycosylation isoforms of FGFR1

Olena Gorbenko1, Galina Ovcharenko, Darija Volkova

  • 1Department of Cell Signalling, Institute of Molecular Biology and Genetics, NAS of Ukraine, Kyiv, Ukraine.

Hybridoma (2005)
|August 12, 2009
PubMed

Insights

Researchers developed new monoclonal antibodies targeting Fibroblast Growth Factor Receptor 1 (FGFR1). These antibodies specifically recognize FGFR1, aiding in the study of its diverse roles in cell functions and diseases.

Area of Science:

  • Molecular Biology
  • Immunology
  • Cell Biology

Background:

  • Fibroblast Growth Factor Receptor 1 (FGFR1) is a receptor tyrosine kinase involved in cell proliferation, differentiation, survival, and tumorigenesis.
  • Multiple FGFR1 isoforms, arising from alternative splicing and post-translational modifications like glycosylation, contribute to its diverse functions.
  • Understanding FGFR1's roles requires specific tools to detect its various forms.

Purpose of the Study:

  • To generate and characterize a panel of monoclonal antibodies specifically targeting Fibroblast Growth Factor Receptor 1 (FGFR1).
  • To utilize these antibodies as research tools for investigating FGFR1 expression and function in biological systems.

Main Methods:

  • Generated monoclonal antibodies using a recombinant FGFR1 extracellular domain fragment (loop II-III) as the antigen.
  • Employed ELISA screening and Western blot analysis to select antibodies recognizing recombinant FGFR1 loop II-III.
  • Validated antibody specificity and recognition of endogenous FGFR1 in 3T3 L1 cells using immunoblot, cyto-, and immunocytochemistry.

Main Results:

  • Successfully isolated a panel of monoclonal antibodies that specifically recognize recombinant FGFR1 loop II-III.
  • Demonstrated that these antibodies can detect endogenous FGFR1 in 3T3 L1 cells, revealing distinct immunoreactive bands potentially representing different isoforms.
  • Confirmed minimal cross-reactivity with FGFR3 and validated FGFR1 detection in cells via various imaging techniques.

Conclusions:

  • The generated monoclonal antibodies are specific tools for detecting Fibroblast Growth Factor Receptor 1 (FGFR1).
  • These antibodies can differentiate between various FGFR1 forms, including potential splice variants and post-translationally modified versions.
  • The antibodies are valuable for studying FGFR1 expression patterns and biological functions in both normal and pathological contexts.

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