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Comparative stability study and aggregate analysis of Bevacizumab marketed formulations using advanced analytical
Arpit Arunkumar Bana1, Nithin Sajeev2, Sabyasachi Halder2
1Department of Pharmaceutical Analysis, Institute of Pharmacy, Nirma University, S. G. Highway, Ahmedabad, 382481, Gujarat, India.
This study characterized two bevacizumab (Bvz) biosimilars in India against the reference product Avastin®. Physicochemical analysis and stress testing revealed minor structural changes, indicating comparable quality and stability for biosimilar evaluation.
Area of Science:
- Biopharmaceutical characterization
- Monoclonal antibody biosimilars
- Oncology therapeutics
Background:
- Bevacizumab (Bvz) is a key biologic therapy in oncology, driving demand for biosimilar development.
- Biosimilar approval requires demonstrating structural and functional similarity to the reference product without clinical significance.
- Comprehensive characterization is crucial for assessing biosimilar quality and stability throughout shelf-life.
Purpose of the Study:
- To compare the physicochemical properties and stability of two Indian bevacizumab (Bvz) biosimilar brands (Bio-1, Bio-2) with the reference product, Avastin®.
- To evaluate potential structural and functional discrepancies that could impact clinical equivalence.
Main Methods:
- Orthogonal physicochemical characterization using techniques like LC-ESI-QTOF, MALDI-TOF, FTIR-ATR, iCIEF, rCE, nrCE, UV280, and RP-HPLC.
- Forced degradation studies under thermal, chemical (pH variations), and mechanical stress conditions.
- Analysis of secondary structure changes via far-UV circular dichroism (CD) and aggregate formation using SEC-MALS.
Main Results:
- Comprehensive physicochemical profiles were established for both biosimilars and the reference product.
- Exposure to elevated temperature and high pH induced minor losses in antiparallel β-sheet conformation (average 15% for Bio-1, 11% for Bio-2).
- Elevated temperature and agitation led to the formation of covalent and non-covalent aggregates, quantified by SEC-MALS.
Conclusions:
- The evaluated bevacizumab biosimilars demonstrated a high degree of similarity to the reference product based on extensive physicochemical characterization.
- Observed minor structural alterations under stress conditions suggest comparable stability profiles, supporting their potential for clinical use.
- The study provides critical data for regulatory assessment of bevacizumab biosimilars marketed in India.
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