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Updated: Aug 23, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Preparation and characterization of anti-human IL-15 monoclonal antibody]
Ping Wang1, Run-tian Wang, Li Wang
1Department of Immunology, Institute of Basic Medical Sciences, Hebei Medical University, Shijiazhuang 050017, China.
Aim:
To prepare monoclonal antibody (mAb) against human interleukin-15 (hIL-15) and identify its characterization.
Methods:
The GST-IL-15 was extracted from the gene-engineering bacteria E. coli and identified by SDS-PAGE. The gel strip containing GST-IL-15 was cut off to immunize BALB/c mice. The splenocytes of immunized mice were fused with Sp2/0 myeloma cells by a routine method and the hybridomas were selected in HAT medium. The hybridoma cells secreting specific antibody were detected by ELISA and cloned by limiting dilution. The stability of the obtained hybridoma cells and the specificity of anti-hIL-15 mAb the hybridoma cells secreted were identified. In addition, the New Zealand rabbits were immunized with the rhIL-15 inclusion body protein (rhIL-15IBP) to prepare the polyclonal antibody (pAb) against hIL-15. A sandwich ELISA was established with the anti-IL-15 mAb and pAb as coating and sandwich antibodies, respectively, to detect hIL-15.
Results:
One hybridoma cell line which could stably secrete specific mAb was obtained. A sandwich ELISA for detecting rhIL-15 protein was established and its sensitivity was as low as 10 microg/L.
Conclusion:
The anti-hIL-15 mAb was prepared successfully. A sandwich ELISA for the detection of hIL-15 was established.
