Development and Validation of a Stability-Indicating Method for Bone Morphogenetic Protein-2

Rev. Sep. Sci. 1(1), 3-16 (2019).

Deblina Biswas, Laure Ray, and Matthew Halquist

Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, VA 23298-0533, USA.


Complex parenteral dosage forms such as subcutaneous implants and liposomes are often associated with diffusion or erosion-based release mechanism of the drug product. One such formulation is INFUSE® Bone Graft. It involves release of a recombinant human bone morphogenetic protein (rhBMP-2) from a biodegradable absorbable collagen sponge (ACS) scaffold. In order to characterize the release of rhBMP-2 from ACS or other scaffolds in a biorelevant media, a simple, precise and accurate stability-indicating high-performance liquid chromatography (HPLC) method was developed and validated. The total run time of the method was 9 min, and the rhBMP-2 retention time was 3.44 min. The dynamic range of the assay was determined to be 0.50–100μμg/mL. The limit of detection (LOD and quantification (LOQ) were observed to be 0.18μμg/mL and 0.50μμg/mL, respectively. Reverse predicted residuals, inter-day accuracy and inter-day precision were all less than 10% including at the LOQ. Forced degradation studies revealed no chromatographic interference from excipients, media or degradant peaks. Mass spectrometric detection was used for further characterization of rhBMP-2 glycoforms in the formulation. Oxidation and deamidation were predicted as the most common degradation mechanism for the protein. The developed method was applied for studying release-testing samples from a novel biorelevant in vitro drug release model (BIVDR). In conclusion, the HPLC- based assay method can be used for studying rhBMP-2 concentrations released from an ACS scaffold in dissolution media.

KEYWORDS: rhBMP-2, RPLC, Stability Indicating Method Validation.

Editor: Dr. Marcello Locatelli, University “G. d’ Annunzio” of Chieti-Pescara, Chieti, Italy.

©2019 Biswas D et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC-BY) which permits any use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

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