Molecular and Biological Characterization of a Prepared Recombinant Human Interferon Alpha 2b Isoform
Shaaban, Rasha; El-Sayed, Wael; Samir, Safia; El-Dabaa, Ehab;
Abstract
Recombinant human interferon alpha2b (rhIFN-α2b) protein is FDA approved for treatment of many tumors and viral diseases. A rhIFN-α2b isoform has been produced and purified from the refolding reaction using high-resolution anion ion exchange chromatography. This isoform has a proper MW (19 kDa) and high purity and homogeneity. The conservation of native linear and conformational epitopes in this isoform was immunologically confirmed by Western blot and ELISA. Mass spectrometry assessment of its intact mass showed average mass (19,337 Da) equivalent to that of the expressed rhIFN-α2b protein without any chemical modification and without the first methionine. Peptide mapping of rhIFN-α2b through tryptic digestion of reductive/alkylated protein using urea as a denaturing agent gave the best pattern. The rhIFN-α2b had a high specific antiviral activity (2.5 × 108 ± 1.1 × 108IU/mg protein). In vivo clearance study of rhIFN-α2b in female SD rats (500 μg/kg, intramuscularly) revealed rapid clearance (elimination half-life 0.54 h with a maximum plasma concentration of 33,792 pg/ml) compared with the commercial rhIFN-α2 (elimination half-life 0.75-0.96 h). In conclusion, the prepared rhIFN-α2b isoform has high purity, homogeneity, native like chemical and structural composition, high antiviral activity, and proper biological stability, which reduce its immunogenicity and raise its therapeutic efficiency.
Other data
Title | Molecular and Biological Characterization of a Prepared Recombinant Human Interferon Alpha 2b Isoform | Authors | Shaaban, Rasha ; El-Sayed, Wael ; Samir, Safia ; El-Dabaa, Ehab | Issue Date | 18-Oct-2018 | Journal | Applied biochemistry and biotechnology | DOI | 1559-0291 10.1007/s12010-018-2908-y |
PubMed ID | 30334171 |
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