Creative Biolabs has established a platform to analyze the peptide map of therapeutic drug candidates, strictly following the ICH Topic Q6b guideline. Utilizing suitable enzymes or chemicals, selective fragmentation of the product is digested into discrete peptides. The resulting peptide fragments are identified by HPLC and other appropriate analytical procedure. The peptide fragments are determined utilizing techniques such as N-terminal sequencing, amino acid compositional analysis, and mass spectrometry. Validated peptide mapping is frequently an appropriate approach to confirm the structure/identity of the desired therapeutic product for lot release purposes.
Comprehensive protein characterization is significant for the quality control of therapeutic protein drugs. It combines a variety of powerful methods that identify and monitor single amino acid changes, modification and degradation products. Peptide mapping is a crucial technique for analyzing the primary structure of proteins. For recombinant therapeutic proteins, peptide mapping is utilized for the initial confirmation of structure characterization. It is also applied for lot-to-lot identity testing in support of manufacturing development and clinical trials. Peptide mapping is utilized as a method of choice for monitoring the genetic stability of recombinant cell lines during bioprocesses. Peptide mapping have recently became much more rapid and convenient, extending its applications for process monitoring and many other fields.
Creative Biolabs provides a comprehensive portfolio of approaches to accurately confirm the protein sequence, identify the modifications, and monitor the routine protein fingerprint for QA/QC, utilizing LC/ESI-MS, Q-TOF systems, liquid chromatography systems and columns, capillary electrophoresis. The protein is cleaved into smaller peptides utilizing a specific protease. The peptides are identified by mass spectrometry and the observed peptides correlate to the amino acid sequence. The confirmed peptides thus determine the specific amino acid sequences, as well as the identity of the protein. N-terminal and C-terminal peptides, amino acid substitution, post translational modification, e.g. glycosylation, disulfide bonds, N-terminal pyroglutamic acid, methionine and tryptophan oxidation can also be confirmed by peptide mapping. Peptide mapping utilizing a single protease will typically result in sequence coverage of 10-90%. Combining two or more proteases this coverage can be enriched to 95-100%. All the proteins are characterized in house in 2-3 weeks.