Elucidation of Structure

At Novabioassays, we provide a suite of analytical services to support the elucidation of structure characterization report for IND or BLA. Our services are listed as in the table.

Attribute Assay Purpose
Primary Structure Mass analysis of intact mAb To verify the molecular mass of the intact mAb
Mass analysis of deglycosylated mAb To verify the molecular mass of deglycosylated mAb
Mass analysis of mass of reduced mAb To verify the primary structure information of the light chain and the heavy chain
Mass analysis of deglycosylated and reduced mAb To verify the primary structure information of the light chain and the deglycosylated heavy chain
Peptide-mapping of mAb and its deglycosylated form with multiple proteases To verify the complete amino acid sequence
To verify the N-terminal sequence of LC and HC
To verify the C-terminal sequence of LC and HC
To determine the identity and estimate the levels of post-translational modifications
N-glycan Profile Peptide-mapping of mAb and its deglycosylated form with multiple proteases To confirm the N-glycosylation site on HC
Released N-glycan analysis from mAb by PNGase F using UPLC-FLD and LC-MS To determine the identity of the glycoforms and their relative abundance
Higher Order Structure Molar Mass and Size Analysis by size-exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) To estimate the molar mass of monomer
Peptide-mapping of mAb for the reduced and non-reduced peptides To confirm the sites of disulfide bond linkage
Thiol assay To determine free sulfhydryl (-SH) content
Circular Dichroism (CD) Compare protein secondary and tertiary structure of two lots
Differential Scanning Calorimetry (DSC) To determine transition temperatures, Tm, as a measure of thermal stability
Quality and Purity SEC-HPLC, iCIEF etc. To evaluate the quality and purity of mAb