Monoclonal Antibody (mAbs) Intact Mass and Subunit Analysis Using Shimadzu Q-TOF LCMS-9030

User Benefits

- Rapid and straightforward workflow for performing mAb characterization using Shimadzu Q-TOF LCMS-9030.

Introduction

Monoclonal Antibody (mAb) therapeutics are a growing group of biopharmaceuticals used to treat medical conditions such as cancer. The characterization of mAb is essential to assess the Critical Quality Attribute (CQA) for product development and regulatory approval. mAbs are large, complex, and heterogeneous proteins which typically require analytical strategies to characterize them to ensure their safety and efficacy as drugs. The use of liquid chromatography coupled with mass spectrometry (LC/MS) is a powerful technique that can be used to characterize of mAb effectively. mAb have a molecular weight of approximately 150 kDa. They are made up of two identical heavy chains, which are each about 50 kDa, and two identical light chains, which are each about 25 kDa. These chains are linked together by disulfide bonds. Each mAb contains a fragment antigen- binding (Fab) region that binds to the antigen and fragment crystallizable (Fc) region at its tail, which contains the N- glycosylation site. Intact mass analysis is a simple and rapid method that can be used to determine the molecular weight of monoclonal antibodies (mAbs). To further characterize mAb, subunit analysis can be performed by cleaving the inter-chain disulfide bonds, which results in the formation of two identical light chains and two identical heavy chains. These subunits can then be analyzed using LCMS to obtain valuable insights into the structural integrity of the mAb protein, which may not be evident from intact mass analysis alone. Subunit analysis can also provide a quick overview of any post-translation modifications (PTMs) or changes in disulfide bonds on the individual light and heavy chains.

December 11, 2023 GMT