Chromatography is the separation of components within a mixture, based on a specific physical molecular attribute. BioPharmaSpec is proficient in a variety of chromatographic techniques and can apply these to your biopharmaceutical product in order to provide quality physicochemical characterization data.
Methodologies and Instrumentation
In reversed Phase-HPLC (RP-HPLC), components are separated based on their hydrophilicity/hydrophobicity. During Ion Exchange chromatography (IEX), separation is achieved through charge differences and with Size Exclusion Chromatography (SEC), separation is based on the size of the components. Hydrophilic interaction chromatography (HILIC) provides separation of analytes based on their polarity.
RP-HPLC is particularly useful for separating deamidated and oxidized peptides from their native counterparts and is often used in peptide mapping experiments to assess these Post Translational Modifications (PTMs).
IEX can be used to purify isoforms within a biopharmaceutical product for further structural analysis and ultimate identification e.g. deamidated forms and monoclonal antibodies with heavy chain C-terminal Lysine.
SEC can be used in conjunction with MS detection to separate components based on their size. UV, RI and Multi Angle Light Scattering (MALS) to also be used alongside SEC to provide an assessment of protein aggregation.
HILIC is particularly useful in separating fluorescently labeled glycans, such as 2-AB, to assess the glycan population.