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Updated: Jun 11, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Production and identification of a monoclonal antibody against human HPPCn]
Jing-rui Huo1, Xi-ning Yao, Ying-fu Liu
1State Key Laboratory of Proteomics, Institute of Radiation Medicine, Academy of Military Medical Science, Beijing 102206, China.
Aim:
To make monoclonal antibody(mAb) against human HPPCn for the use in research on HPPCn's function and its relationship with liver diseases.
Methods:
The female BALB/c mice were immunized with the recombinant HPPCn proteins. Splenocytes and Sp2/0 cells were fused with PEG-1500. The positive clone was identified through indirect ELISA and then subcloned by limited dilution. Indirect ELISA, Western blot and Ig sub-class identification kit were used to identify the mAb's properties. By immunofluorescence experiments, we studied the cellular localization of HPPCn. The mAb epitope was also analyzed using peptide phage display technology.
Results:
An anti-HPPCn mAb, named W2-D5, was obtained. It belongs to IgG1 subclass. It could specially bind to human HPPCn. Furthermore, by immunofluorescence results, wo confirmed HPPCn located in the nucleus and our mAb could combined with the natural protein. With the mAb, the minimal detectable concentration was 0.1 μg/L for HPPCn; The peptide sequence of HPPCn₇₋₁₃;(IHLELRN)was identified as the epitope of the mAb.
Conclusion:
An anti-HPPCn mAb with high specificity and high affinity was successfully obtained.
