Protein Identification & Characterization Services

Structural, Physicochemical and Biophysical Characterization Methods

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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

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

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