Monoclonal antibodies to a monkeypox virus polypeptide determinant

Area of Science:

• Immunology
• Virology
• Molecular Biology

Background:

• Monkeypox virus (MPV) is a significant public health concern.
• Accurate diagnostic tools and understanding of viral antigens are crucial for MPV control.

Purpose of the Study:

• To characterize novel murine monoclonal antibodies against MPV.
• To identify specific MPV antigens recognized by these antibodies.
• To assess the specificity and neutralizing capacity of the developed antibodies.

Main Methods:

• Characterization of immunoglobulin G1 (IgG1) murine monoclones.
• Indirect fluorescent-antibody test (IFAT) and enzyme immunoassay (EIA) for antibody reactivity.
• Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) transblot assay to identify target polypeptide.
• Adsorption assay using vaccinia virus-infected cells.
• Cross-reactivity testing against related orthopoxviruses (cowpox, variola, vaccinia).

Main Results:

• Three MPV-specific murine monoclonal antibodies were identified, all of the IgG1 isotype.
• Antibodies showed strong reactivity in IFAT (4+) and EIA.
• Monoclonal antibodies recognized a 15,500-molecular-weight MPV polypeptide.
• Reactivity was specific to MPV and did not cross-react with cowpox, variola, or vaccinia viruses.
• The antibodies did not neutralize MPV infectivity.

Conclusions:

• The characterized monoclonal antibodies are specific to MPV and target a distinct 15.5 kDa polypeptide.
• These antibodies hold potential for use in developing specific diagnostic assays for monkeypox virus detection.
• Lack of neutralization suggests their utility in diagnostic rather than therapeutic applications.