Preparation of Polyclonal Antibodies Based on Inhibin α Subunits of Ferritin Nanocarriers
In order to prepare a polyclonal antibody against inhibin α(INH-α)with high potency and multi-animal applicability,spe-cies homology of INH-α was analyzed by bioinformatics,and pET-32α was used as the backbone for the construction of INH-α and INH-α-Ferritin expression vectors,and the recombinant proteins were used for immunization of Japanese lop-eared rabbits after pro-karyotic expression of purified recombinant proteins,the titer of antibody in rabbit serum was determined by indirect ELISA.The re-sults showed that the INH-α and INH-α-Ferritin expression vectors were successfully constructed,and the recombinant proteins in the form of 35 kDa and 50 kDa inclusion bodies were obtained in 0.4 mmol/L IPTG at 37 ℃ for 6 h,respectively.A high titer rabbit poly-clonal antibody was obtained by immunization,the titer of antiserum of INH-α-Ferritin reached 1∶19 531 250.Bioinformatics analysis showed that INH-α was highly homologous and conserved among mammals.In this study,we successfully constructed INH-α and INH-α-Ferritin prokaryotic expression vectors,and significantly improved the potency of polyclonal antibodies by INH-α-Ferritin,which lays the foundation for the study of improving the fertility of mammalian livestock.
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