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Human single chain monoclonal antibody that recognizes matrix protein of heterologous influenza A virus subtypes

Ornnuthchar Poungpair1, Wanpen Chaicumpa, Kasem Kulkeaw

  • 1Graduate Program, Faculty of Allied Health Science, Thammasat University, Pathumthani 12120, Thailand.

Journal of Virological Methods
|May 16, 2009
PubMed

Insights

Researchers developed human single monoclonal antibody fragments (HuScFvs) targeting the influenza A virus matrix protein (M1). One antibody fragment blocked M1-RNA binding, showing potential as a broad-spectrum antiviral therapy.

Area of Science:

  • Virology
  • Immunology
  • Biotechnology

Background:

  • The influenza A virus matrix protein (M1) is crucial for viral replication and assembly.
  • M1 is a promising target for antiviral therapies, including drugs, siRNA, and antibodies.
  • No therapeutic antibody has been developed to inhibit M1's multifaceted functions.

Purpose of the Study:

  • To generate human single monoclonal antibody fragments (HuScFvs) against the influenza A virus M1 protein.
  • To evaluate the potential of these HuScFvs as a novel antiviral strategy against influenza A.
  • To investigate the diversity of epitopes and affinities of the generated HuScFvs.

Main Methods:

  • Production of full-length recombinant M1 (rM1) from highly pathogenic avian influenza virus A/H5N1.
  • Phage bio-panning using a human antibody phage display library against rM1.
  • Restriction Fragment Length Polymorphism (RFLP) analysis to assess HuScFv diversity.
  • Enzyme-Linked Immunosorbent Assay (ELISA), immunostaining, and immunofluorescence to test HuScFv binding to native M1 in infected cells.

Main Results:

  • Several phage clones displaying HuScFvs that bind to rM1 were successfully isolated.
  • RFLP analysis indicated epitope and affinity diversity among the selected HuScFvs.
  • One specific HuScFv demonstrated the ability to block the binding of M1 protein to viral RNA.
  • Tested HuScFvs showed binding to native M1 from homologous and heterologous influenza A viruses.

Conclusions:

  • Human single monoclonal antibody fragments (HuScFvs) against influenza A virus M1 protein were successfully generated.
  • A specific HuScFv was identified that inhibits a key M1 function (RNA binding).
  • Development of cell-penetrating HuScFvs holds promise for a broad-spectrum anti-influenza A therapeutic strategy.

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