Monoclonal antibody AP-282 recognizes a marker for human polymorphonuclear granules

Area of Science:

• Immunology
• Cell Biology
• Hematology

Background:

• Hybridoma technology enables the production of specific monoclonal antibodies (MAbs).
• Human neutrophils play critical roles in innate immunity and inflammation.
• Characterizing specific cell markers is essential for diagnostic and research applications.

Purpose of the Study:

• To develop and characterize a novel monoclonal antibody (MAb) for identifying human neutrophils.
• To investigate the specificity and utility of MAb AP-282 as a cellular marker.

Main Methods:

• Production of hybridoma AP-282 by fusing mouse plasmacytoma cells with splenocytes.
• Immunological reactivity testing of MAb AP-282 against various human blood and bone marrow cells.
• Antigen characterization using fixation resistance and APAAP staining techniques.
• Comparative analysis with known anti-granulocyte antibodies (MPO, elastase, cathepsin G, lactoferrin).

Main Results:

• MAb AP-282, a mouse IgG1, strongly reacts with neutrophils and their bone marrow precursors.
• The antibody shows weak reactivity with monocytes and no reactivity with eosinophils.
• The target antigen is resistant to formalin but sensitive to acetic acid fixation.
• AP-282 specifically labels neutrophils in various tissue preparations via APAAP staining.
• The antigen is distinct from myeloperoxidase, elastase, cathepsin G, and lactoferrin.

Conclusions:

• MAb AP-282 identifies a novel cytoplasmic antigen specific to neutrophils.
• This antibody serves as a valuable new marker for neutrophil identification in research and potentially diagnostics.