Generation of Novel Monoclonal Antibodies Recognizing Rabbit CD34 Antigen.

Area of Science:

• Immunology
• Hematology
• Stem Cell Biology

Background:

• Rabbits are vital experimental models in translational research and stem cell therapy.
• Identifying rabbit hematopoietic stem/progenitor cells is challenging due to the lack of specific CD34 antibodies.
• CD34 is a standard marker for hematopoietic stem/progenitor cells in humans and other species.

Purpose of the Study:

• To develop novel monoclonal antibodies (mAbs) highly specific to the rabbit CD34 antigen.
• To overcome the limitation in studying rabbit hematopoietic stem cells.

Main Methods:

• Hybridoma technology utilizing synthetic peptides and recombinant protein for immunization.
• Screening of antibody binding activity and specificity via ELISA, flow cytometry, and Western blot.
• Selection and purification of four specific anti-rabbit CD34 mAbs.

Main Results:

• Four novel mAbs (58/47/26, 58/47/34, 182/7/80, and 575/36/8) were successfully generated and purified.
• The mAbs specifically recognize CD34 on rabbit bone marrow and peripheral blood cells.
• Detected CD34+ cells (2-3%) and CD45+ cells (approx. 2%) in rabbit bone marrow, indicating primitive hematopoietic stem cells and progenitors.

Conclusions:

• The developed mAbs are specific tools for identifying rabbit CD34+ cells.
• These antibodies will advance research in rabbit hematopoiesis and stem cell transplantation models.
• Enables new immunological applications and enhances rabbit models for regenerative medicine.