Can CE-MS be used for Antibody Quantitation in Biological Samples?
Investigator, Bioanalysis, Immunogenicity & Biomarkers, IVIVT GSK
Current liquid chromatography-mass spectrometry (LC-MS) methods used to detect monoclonal antibodies (mAbs) in biological samples are focused on targeted tryptic peptide quantitation, which requires that the mAbs undergo tryptic digestion before analysis. The use of digestion complicates the workflow and can affect the results obtained. Therefore, there is an increasing interest to simplify the workflow, by developing improved methods which can detect the intact antibody or larger antibody fragments.
In this presentation, we describe how a capillary electrophoresis-mass spectometry (CE-MS) method has been developed to quantitate a reduced monoclonal antibody in biological samples.
We will highlight the factors you need to consider when developing a CE-MS method to detect antibody fragments of 25 and 50k Da, and show how injection and separation conditions can affect the sensitivity and peak resolution obtained.
Finally, we will demonstrate how using a surrogate mAb improves the reproducibility of electro-kinetic injections and highlights how CE-MS is now capable of detecting a mAb at low ng/mL sensitivities.
In this webinar we will be discussing:
Why CESI-MS is ideal for detecting intact proteins
How CESI-MS can detect low concentrations (ng/mL) of mAbs in biological samples
What factors you need to consider when developing a CE-MS method to quantitate intact proteins