Date of Publication :25th March 2024

Abstract:Antibodies generated by a unique clone of B cells are called monoclonal antibodies, and mAbs from the exact B cell clone bind to the identical epitope(s). In diagnostics and therapeutics, mAbs serve as an excellent tool. NDV is a highly contagious pathogen causing nervous, respiratory, and enteric diseases amongst birds worldwide. Since poultry farming is done chiefly in rural areas, developing cheap, easy, and effective diagnostic kits, such as rapid antigen detection kits, can be helpful in the quick and easy detection of NDV infection. Therefore, this study is predicted on generating the NDV-specific hybridoma that can be used to produce mAbs suitable for developing rapid antigen detection kits. For generating the NDV- specific B cells, three mice were immunized subcutaneously with NDV after getting the required permission from Institutional Animal Ethics Committee (IAEC, SSBS, SIU). After the secondary and booster immunization, hemagglutinin inhibition assay (HIA) was performed with NDV specific antibodies and serum of experimental animals. The mouse expressing the highest antibody titer, primed with NDV immunization four days before sacrificing it. Splenocytes were prepared from sacrificed mice spleen and fused with SP2/0 myeloma cells using PEG and electrical pulse. The fused hybridoma cells were grown on hypoxanthine- aminopterin - thymidine (HAT) media. Cultured cells HIA supernatant was utilized for detection of NDV-specific antibodies. Two out of four cultures were positive for the particular antibody and it was observed that, IgG level was elevated in the serum whereas, IgM + IgA were significantly elevated in the supernatant. Hybridoma producing anti-NDV specific antibodies have been generated. Successful expansion of NDV specific hybridoma clones will lead to the development of monoclonal antibodies that can be used for making the lateral flow assay kit suitable for rapid virus detection.

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