Figures & data
Figure 1. Comparison of HPLC peptide mapping between the etanercept reference product (Enbrel®) and its biosimilars. (B) was the partial zoom of (A) (26–35min). Line 1 to 7 was blank control, biosimilar 1 (Lot 1), biosimilar 1 (Lot 2), biosimilar 1 (Lot 3), biosimilar 1 (Lot 4), reference product (Wyeth, Lot F12508), and biosimilar 2 (Lot 1), respectively.
Figure 2A and B. Comparison of deconvoluted masses of Fc (A and B).
Figure 2C and D. Comparison of reduced Fc (C and D) between the reference product and two etanercept biosimilar products.
Figure 3. Mirror plots of LC-MS peptide maps from 4 h tryptic digests of the Fc of the reference product and two etanercept biosimilar products. (A) Comparison of LC-MS (TIC) chromatogram between the reference and biosimilar 1 products; (B) Comparison of LC-MS (TIC) chromatogram between the reference and biosimilar 2 products. (C) A zoom charge-reduced, isotope-deconvoluted LC-MS chromatograms from 22–36 min.
Figure 4. Mirror plots of zoom charge-reduced, isotope-deconvoluted LC-MS chromatograms (1–6min) of the FC of reference and two etanercept biosimilar products. (A) Comparison of biosimilar 1 with the reference product; (B) Comparison of biosimilar 2 with the reference product.
Figure 5. Charge-reduced, isotope-deconvoluted MSE spectra (fragment ion matched profiles) of tryptic peptide T16. (A) T16 (EEMTK) of biosimilar 1; (B) T16 (DELTK) of biosimilar 2.
Figure 6. Non-reduced SDS-PAGE analysis of the reference etanercept product and its biosimilar products before (A) and after (B) desialylation. E1 is the reference product (Enbrel® Lot F12508); Y1, Y2, Y3 and Y4 are 4 lots of biosimilar 1. Q1 is one commercial lot of biosimilar 2.
Table 1. TNFα receptor II–Fc fusion proteins*
Figure 7. Comparison of IEX charge variant between the reference etanercept product and its biosimilar products (after desialylation). E1 is the reference product (Enbrel® Lot F12508); Y1, Y2, Y3 and Y4 are 4 lots of biosimilar 1. Q1 is one commercial lot of biosimilar 2.
Figure 8. Typical CE-LIF electropherogram of released N-oligosaccharides from etanercept reference and biosimilar products before (A) and after desialylation (B). E1 is the reference product (Enbrel® Lot F12508); Y1, Y2, Y3 and Y4 are 4 lots of biosimilar 1. Q1 is one commercial lot of biosimilar 2.
Table 2. Composition percentage of N- Glycans from the innovator Enbrel® and two commercially available biosimilar products
Table 3. Composition percentage of N-oligosaccharides from desialylated TNFα receptor II–Fc fusion proteins*
Table 4. Comparison of kinetics fitting results between the etanercept reference product and two commercially available biosimilars*
Figure 9. Four-parameters fitting curve for the potency determination of the etanercept reference and biosimilar products. E1 is the reference product (Enbrel® Lot F12508); Y1, Y2, Y3 and Y4 are 4 lots of biosimilar 1. Q1 is one commercial lot of biosimilar 2.
