BioPharmaSpec provides a comprehensive range of protein characterization services to meet global regulatory requirements for biopharmaceutical characterization, whether you are developing a biosimilar, a novel molecule, a peptide, an antibody, a bi/multi-specific, a drug-conjugate or any other molecule!
The harmonized ICH guideline Q6B suggests that characterization of a biopharmaceutical should include:
Structural & Physicochemical Characterization
Biophysical Characterization
Purity and Impurities
Host Cell Protein (HCP) Analysis & Detection
BioPharmaSpec has developed a service for MS based HCP detection and quantitation as an orthogonal support to HCP-ELISA.
Process Related Impurities
BioPharmaSpec provides methods for the analysis of a significant range of process related impurities. These methods have been developed to allow identification and quantitation to low ppm/ppb limits.
During biopharmaceutical development, detailed and thorough characterization should be performed. If significant process changes are made, the guidelines state that characterization should also be performed following these changes, in order to prove that there have been no major changes to the structure or function of your product.
It is important to make use of the most up-to-date protein characterization methods. The guidelines advise that New analytical technology and modifications to existing technology are continually being developed and should be utilized when appropriate.
BioPharmaSpec uses state-of-the-art instrumentation and a pioneering approach to methodology to characterize your biopharmaceutical to the highest technical standard.
Protein Characterization Methods
| Amino Acid Analysis | Triplicate hydrolysis, duplicate injection |
| N-terminal Sequencing | 15 cycles (of light and heavy chains) |
| C-terminal sequencing | Determined through molecular weight and peptide mapping |
| Peptide Mapping | At least three digestion strategies LC/ES-MS, Q-TOF |
| Glycosylation site occupancy | Assessment of the degree of glycosylation site occupancy |
| Primary sequence confirmation | MS based sequencing and Ile/Leu differentiation using Edman for the full sequence |
| Disulfide Bridge Assessment | Determined through peptide mapping |
| Monosaccharide Composition Analysis | Neutral and amino sugars Sialic acids |
| Oligosaccharide profiling | HILIC analysis of 2-AB labelled oligosaccharides covering the analysis of total N-glycans Linkage (methylation) analysis |
| Intact molecular weight analysis | As received, following reduction, following de-N-glycosylation, following reduction and de-N-glycosylation. On line RP-HPLC-MS |
| icIEF | With and without Carboxypeptidase B treatment |
| CE-SDS | Non reducing and reducing |
| RP-HPLC with UV detection | Performed on the sample as received |
| Ion Exchange Chromatography | Isoforms can be collected and characterized |
| DSC | To assess melting temperature |
| CD | For far-UV (for secondary structure) and near UV (for tertiary structure) |
| FT-IR | Secondary structure |
| IF | Secondary and tertiary structure |
| 1D and 2D NMR | Secondary and tertiary structure |
| SEC-MALS | Aggregation assessment |
| SV-AUC | Orthogonal aggregation assessment |
| Cell based assay(s) | Functional assessment |
| Binding assay(s) | Assessment of binding affinity to receptors, ligands etc |
The Role of Protein Characterization in Product Development
The above protein characterization methods can be included in specific studies in order to support key development decisions. These characterization techniques and the expert data interpretation and technical summaries you will receive from BioPharmaSpec scientists, will help you choose the best cell clone, provide batch-to-batch comparisons or make innovator to biosimilar comparisons. We support activities such as:
- New, innovative product characterization
- Structural and physicochemical characterization to ICH Q6B
- Comparability studies
- Biosimilarity studies
- Forced degradation studies
- Stability studies
- Product impurity analysis and quantitation
- Host Cell Protein detection using Mass Spectrometry
Need protein structure characterization services?
Work with BioPharmaSpec scientists to design effective characterization strategies that will allow you to make solid development decisions based on quality analytical data.