Icosagen has a dedicated department for protein analytics to ensure meticulous product characterization for optimal quality of both antibodies and recombinant proteins.
For customized solutions we have a comprehensive list of analytical options available to cover all the needs for proteomic characterization.
Involves standard methods to analyse molecular weight, check molecule expression and secretion during production as well as confirm that produced antibodies are functional and bind to the target antigen.
Analytics play a key role in protein production in order to get functional proteins with high purity. Most commonly used methods are:
Coomassie SDS-PAGE and LabChip capillary SDS — a standard test to analyse:
molecular weight
check molecule expression and secretion during production
estimate production purity during purification.
Western Blot and ELISA test to:
detect produced proteins from the sample
confirm that produced antibodies are functional and bind to the target antigen.
Additional: Octet Red96 — for measuring protein concentration in culture media.
Final product quality control
Product quantitation and molecular weight sizing in addition to impurity and homogeneity analysis. Determine charge distribution as a part of modifications analysis.
Analytical Size-Exclusion Chromatography (aSEC) — used to:
characterize final purified or thawed fraction by molecular weight to monitor the purity of the product and detect the presence of aggregates.
Analytical Ion-Exchange chromatography (aIEX) — used to:
determine protein charge distribution as a part of modifications analysis
control final fraction of bi-specific antibodies to assure no monospecific antibody versions are present, in case there is sufficient charge difference between these two. This is usually supplemented with mass-spectrometry.
LAL-test — used to:
determine bacterial endotoxins in purified recombinant protein samples.
LabChip GXII Protein Characterization System — provides the complete solution for product quantitation and molecular weight sizing or impurity and homogeneity analysis
Product characterization
For customized solutions we have a comprehensive list of analytical options for glycosylation pattern analysis, stability studies in different stress conditions as well as formulation studies.
All-in-one protein stability analysis with the Uncle — a stability platform that uses fluorescence, static light scattering (SLS) and dynamic light scattering (DLS) detection methods to characterize protein stability. Allows to:
rank the stability of your products based on thermal melting and aggregation (Tm and Tagg)
compare different formulations to find the most favorable conditions
find excipients that prevent or delay the onset of aggregation
identify the presence of aggregates or impurities
confirm that your molecule is the size you expect it to be and
isothermal stability
Octet Red96 — is a biolayer interferometry biosensor for:
measuring protein concentration (in the culture media)
kinetics and affinity of intermolecular interactions
assessing binding specificity
epitope mapping
rates of association and dissociation
Glycosylation pattern analysis:
Hydrophilic interaction liquid chromatography (HILIC) — an alternative high-performance liquid chromatography (HPLC) that can be used for determination of N-glycans or fucosylation levels in antibodies.
LabChip Glycosylation pattern analysis — the microchip-based CE analysis to determine the distribution peaks for further analysis of relative abundance of the major N-glycan types found on antibodies, which contain primarily neutral N-linked glycans consisting of complex, hybrids, and high-mannose type oligosaccharides.
Isoelectric focusing (IEF) and charge variant analysis with LabChip