Reversed phase high performance liquid chromatography for monitoring type-16 human papilloma virus like particles

Abstract

Quantification of virus like particles (VLPs) is challenging due to the large size and structural heterogeneity of the molecule. Structural heterogeneity arises due to random assemblage of L1 protein with other host cell proteins. With multiple VLPs under development as potential biotherapeutic products, defining their identity, purity and potency is of great interest. Most analytical methods used at present for VLP quantification are laborious and at best semi-quantitative. Thus, there is a need for an analytical technique that is capable of quantitatively estimating both purity and titre at all stages of the process. In this study, we propose use of Reversed phase chromatography for separation and identification of VLP L1 proteins in inhouse crude and purified samplesand quantification of L1 protein at different process points. The proposed method takes about 20 min and has been demonstrated to be highly sensitive, robust, and selective as confirmed through mass spectrometry.