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Related Experiment Videos

Two spatially distant epitopes of human lactoferrin

D Caccavo1, A Afeltra, F Guido

  • 1Department of Medicina Clinica, University La Sapienza, Rome, Italy.

Hybridoma
|August 1, 1996
PubMed
Summary

Two new monoclonal antibodies (MAbs) targeting lactoferrin (LF) were developed. These MAbs are valuable tools for studying autoantibodies to LF and measuring soluble LF, a marker of neutrophil activation.

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Area of Science:

  • Immunology
  • Biochemistry

Background:

  • Lactoferrin (LF) is an iron-binding protein implicated as a potential target for autoantibodies in autoimmune diseases like rheumatoid arthritis and inflammatory bowel diseases.
  • Antineutrophil cytoplasmic antibodies (ANCA) are associated with several autoimmune conditions, and LF is a candidate autoantigen.

Purpose of the Study:

  • To produce and characterize novel monoclonal antibodies (MAbs) against human lactoferrin (LF).
  • To evaluate the utility of these MAbs in detecting LF and its role in neutrophil activation.

Main Methods:

  • Production and characterization of two mouse monoclonal antibodies (MAbs) against human LF (AGM 10.14 and AGM 2.29).
  • Epitope mapping using cross-blocking experiments.
  • Indirect immunofluorescence on ethanol-fixed neutrophils.

Related Experiment Videos

  • Development of a double determinant immunoassay (DDIA) using the MAbs to quantify soluble LF.

    • Main Results:

    • Two MAbs, AGM 10.14 (IgG1) and AGM 2.29 (IgG2b), were generated, recognizing distinct epitopes on LF.
    • Both MAbs showed perinuclear staining on neutrophils via indirect immunofluorescence.
    • A DDIA utilizing these MAbs successfully quantified soluble LF in a range of 12–780 ng/ml, demonstrating high sensitivity.

    Conclusions:

    • The developed MAbs (AGM 10.14 and AGM 2.29) are specific reagents for human LF.
    • These MAbs are useful for investigating the role of anti-LF autoantibodies in autoimmune diseases.
    • The DDIA assay provides a sensitive method for measuring soluble LF, a marker of neutrophil activation.