Research Article: Quality by Design–Based Assessment for Analytical Similarity of Adalimumab Biosimilar HLX03 to Humira®

Date Published: May 8, 2020

Publisher: Springer International Publishing

Author(s): Erhui Zhang, Liqi Xie, Peilan Qin, Lihong Lu, Yanpeng Xu, Wenyuan Gao, Linlin Wang, Michael Hongwei Xie, Weidong Jiang, Scott Liu.


Quality by design (QbD) is an efficient but challenging approach for the development of biosimilar due to the complex relationship among process, quality, and efficacy. Here, the analytical similarity of adalimumab biosimilar HLX03 to Humira® was successfully established following a QbD quality study. Quality target product profile (QTPP) of HLX03 was first generated according to the public available information and initial characterization of 3 batches of Humira®. The critical quality attributes (CQAs) were then identified through risk assessment according to impact of each quality attribute on efficacy and safety. The anticipated range for each CQA was derived from similarity acceptance range and/or the corresponding regulatory guidelines. Finally, a panel of advanced and orthogonal physicochemical and functional tests and comparison of 6 batches of HLX03 and 10 batches of the reference standard demonstrated high similarity of HLX03 to Humira®, except for slightly lower percentage of high mannosylated glycans (%HM) in HLX03 which had no effect on FcγRIII binding and antibody-dependent cell-mediated cytotoxicity (ADCC) activity in human peripheral blood mononuclear cell (PBMC). All above demonstrated the feasibility and efficiency of QbD-based similarity assessment of a biosimilar monoclonal antibody (mAb).

Partial Text

Development of biosimilars will lead to more affordable biological treatments and increase patient access to otherwise expensive therapies. It is especially important for therapeutic mAbs, which represents a major cost burden for health-care providers on rapidly increasing usage (1). Because of the complexity of antibody structure as well as its sensitive nature to the manufacturing process, the development of biosimilar faces great technical challenges and risks of not being similar.

According to NMPA, EMA, FDA, and ICH regulatory guidelines for the development of biosimilar products (3,11,16–18), a stepwise approach incorporating QbD was used to evaluate the similarity of HLX03 to CN-Humira®. A total of 6 batches of HLX03 and 10 batches of CN-Humira® were characterized to compare their primary structure, higher-order structure, charge heterogeneity and post-translational modifications, glycosylation, molecular size heterogeneity, immunological properties, biological activity, as well as forced degradation behavior by a series of state-of-the-art and orthogonal analytical methods. The analytical similarity study was performed following a QbD approach, from establishment of QTPP, selection of analytical technologies and methods, risk assessments to determine CQAs, to the establishment of acceptance criteria for the assessment of analytical similarity.

QbD has become an integral part of therapeutic protein development to obtain high and constant product quality with less risk of batch failures. However, effective implementation of QbD is still a challenge in the biopharmaceutical industries due to complex relationships among quality, efficacy, and process procedures. Here, the similarity assessment of adalimumab biosimilar HLX03 to CN-Humira® demonstrated an example of QbD-based analytical similarity study for biosimilar development.

Physicochemical and functional tests demonstrated high similarity of HLX03 to CN-Humira®, illustrating an example of QbD-based analytical similarity assessment for a biosimilar mAb development.