Development of a novel and rapid HPLC-based method for a comparative assessment of biosimilar structural heterogeneity and biological activity

Authors:: Poster Author(s)

Jackie Cullinan FDA/CDER (Student), Rachel CarleyFDA/CDER, Talia Faison FDA/CDER, Sarah Johnson FDA/ CDER, Cyrus Agarabi FDA/CDER, Scott Lute FDA/CDER (Mentor)

Center:: Contributing Office

Center for Drug Evaluation and Research

Abstract

Poster Abstract

Monoclonal antibody (mAb) drug products are the largest product class of US-licensed biosimilars. The glycosylation post-translational modifications (PTMs) on the Fc effector region of mAbs are a well-defined critical quality attribute (CQA) with significant impact on the Fc effector mediated antibody-dependent cellular cytotoxicity (ADCC)mechanism of action (MoA) for many of the mAb drug products. Established methods to assess glycan structures and Fc effector activity are costly, time-consuming, require in-depth sample preparation and highly skilled analysts. Implementation of rapid and high throughput methods may allow for faster development of biosimilar drug products. Recently, an affinity chromatography based technology, the TSKgel FcR-IIIA-NPR HPLC affinity column, emerged claiming to provide a faster and straight-forward approach to assessing mAb biosimilar glycosylation and Fc effector mediated activity. We propose to ascertain the feasibility of the TSKgel FcR-IIIA-NPR HPLC affinity column as an rapid assay to identify differences in glycosylation PTMs between proposed biosimilars and their reference products. TOSOH claims the use of TSKgel FcR-IIIA-NPR HPLC affinity column will result in a chromatogram with 3 peaks correlating to high, mid, and low affinity to the column, thereby identifying differences in glycosylation patterns between drug products and their proposed biosimilars faster than traditional methods. In order to support this claim, first a method was established with the well-characterized NIST mAb. which will include refining of HPLC method conditions e.g. mobile phase buffer conditions until reproducible separation of glycan fractions is achieved with the NIST mAb. Future “proof-of-concept" will be demonstrated through application of the method to the reference drug product infliximab and its three biosimilars: infliximab-dyyb, infliximab-abda, and infliximab-axxq. Lastly, the specific glycan content of each HPLC peak will be confirmed via established mass spectrometry (MS) methodologies for NIST mAb, infliximab, and its biosimilars.