Figure 7. Proteins identified in each band of SDS-PAGE gel
resolved proteins by LC-MS/MS.
Figure 8. EpiMatrix ClustiMer immunogenicity scale. Clustimers
derived from residual proteins in API vary in their calculated
immunogenicity risk. Clustimers with low human homology and their scores
were identified and calculated in the EpiMatrix program. Clustimers were
binned according to their score and are displayed on the right side of
the scale. Number in parenthesis indicates multiple peptides from a
given protein are included in the range. Foreign proteins and human CLIP
are shown on the left side of the scale to give relative risk.
Immunogenicity risk increases from green (minimal) to red (high).
Abbreviations: HCV= Hepatitis C virus, CLIP= Class II-associated
invariant chain peptide. EBV=Epstein-Barr virus. TRPCIF=Transcriptional
Regulatory Protein CpxR IcIR Family, AHRC= Alkyl Hydroperoxide Reductase
C, 30SRPS10= 30S Ribosomal Protein S10, CspC= Cold Shock-like Protein C.
Figure 9. MK-1454 API isolated and purified with the new process
resolved on SDS-PAGE with silver stain (a), Coomassie blue stain (b).BSA, bovine serum albumin, MK-1454 157, 165, 166 are three small scale
biocatalytic API isolated with the new workup procedure. C-10 mg/mL,
represents a chemically synthesized MK-1454 control that is free of
protein impurities.