
Biophysical characterization of biologics is critical in the development of biopharmaceuticals such as innovator biologics and biosimilars. Many aspects of biologic drug function, activity, and stability are affected by the physical and biological behavior of proteins. Biophysical analytical techniques, for example, can aid in monitoring or confirming conformational integrity, the nature of the protein’s folded state, and how peptides interact to form complex physical structures. Consideration of specific interactions between drug substance and excipients that might potentially alter protein structure and product stability, is critical during formulation development, stability studies, and comparability. Protein degradation and aggregation can also be studied using biophysical techniques.
Aragen has established an advanced state-of-the-art, fully equipped analytical laboratory with highly qualified analytical scientists to perform a comprehensive characterization of biologics.
Analytics in Drug Discovery and Development Process using array of Instruments.
Protein Analytics | Quality Attribute | Platform |
---|---|---|
Intact Mass (non-reduced, reduced, +/-PNGaseF) | Primary Structure – MW | LC – MS /QTOF (Agilent) |
PepMap | Identity an Primary Structure PTMs | LC – MS /QTOF (Agilent) |
PepMap UV | Identity | LC – MS /QTOF (Agilent) |
N-Glycan profiling by RP-MS or HILIC-MS | Primary Structure – PTMs | LC – MS /QTOF (Agilent) |
Glycation | Primary Structure – PTMs | LC – MS /QTOF (Agilent) |
Absolute mass for monomer multimers, and aggregate | Purity – Size variants | SEC – HPLC – MALS (Wyatt) |
Tm, Tagg, Size and PDI | Thermal Stability, Higher Order Structure (HOS) | SLS /DLS (Unchained labs) |
Sialic Acid Content | Primary Structure – PTMs | UPLC (Agilent) |
N -Glycan profiling | Primary Structure – PTMs | HILIC -FLD /UPLC (Agilent) |
Charge variants | Purity – Charge variants | IEX/HPLC (Agilent) |
Size variants (aggregates ) | Purity – Size variants | SEC-HPLC (Agilent) |
Hydrophobicity variants | Purity – Protein related substances and impurities | RP or HIC/UPLC (Agilent) |
Glycation | Primary Structure – PTMs | RP or HIC/UPLC (Agilent) |
Protein concentration by Protein-ARP | Titer/Quantitation | BAC – HPLC (Agilent) |
Free – thiol | Primary Structure – Free thiol | FLD – UPLC (Agilent) |
Protein concentration by A280 | Quantity | UV – VIS |
A280, A320, Appearance | General – Appearance, Trubidity | UV – VIS |
Free-thiol by Ellman’s assay | Primary Structure – Free thiol | UV – VIS |
Protein Analytics | Quality Attribute | Platform |
---|---|---|
N – Glycan profiling | Primary Structure -PTMs | CE – SDS (SCIEX) |
Charge variants | Purity | cIEF(SCIEX) |
Charge variants | Purity | CZE (SCIEX) |
Slab gel – charge variants | Purity | IEF, Gel Electrophoresis |
Size variants | Purity | SDS-PAGE, Gel Electrophoresis |
Western blot | Identity | Gel Electrophoresis |
Binding kinetics (kon, koff, Kd) | Potency, Identity | BLI (Sartorious /ForteBio) or SPR (Cytiva /Biacore) |
FCR and C1Q screening | Potency, Half – life, Safety | BLI (Sartorious /ForteBio) or SPR (Cytiva /Biacore |
Protein/antigen titer | Quantity | BLI (Sartorious /ForteBio) |
Epitope binning | Epitope characterization | BLI (Sartorious /ForteBio) or SPR (Cytiva /Biacore) |
Functional blocking | Potency, Identity | BLI (Sartorious /ForteBio) or SPR (Cytiva /Biacore) |
Yes /No binding | Identity | BLI (Sartorious /ForteBio) or SPR (Cytiva /Biacore) |
Binding | Potency | SPR (Cytiva /Biacore) |
Host Cell Proteins (Cygnus) | Safety | ELISA /Multimode plate reader (SpectraMax) |
Residual Protein A/G/L | Safety | ELISA /Multimode plate reader (SpectraMax) |
Endotoxin Testing | Safety | LAL /Charles River EndoSafe |
Bioburden, Sterility Test | Safety | Culturing |
Comprehensive Biologics Characterization
Instrumentation
For characterizing antibody epitopes and multiprotein complexes of biological significance – detect presence of specific proteins with minimal interference from complex matrices.
Key Features
Case Study : Octet-based FcR and C1q screen
Agilent 1290 UPLC + 6530B QToF
Case study : Sequence of target molecule – peptide mapping
Case Study : Determination of the sugar content of bioreactor antibodies – N-glycan analysis
Case study : Assessment and rigorous testing of the quality of purified target molecules
Case study : Determining the sequence of target molecule – peptide mapping
Formulation Development using DLS/Uncle
Key Features
Measuring various parameters on multiple samples simultaneously using Uncle
Developability Process to obtain Insight into Structure/Function relationships and potential Critical Quality Attributes (pCQAs)
Characterization by Multimode Plate Readers
Combining scientific expertise with expedited protocols to deliver target-specific antibody solutions.
Solutions for the stable expression and purification of novel molecules.