Development of a capillary zone electrophoresis method for dose determination in a tetravalent dengue vaccine candidate

Abstract

Dengue is known to cause morbidity and mortality worldwide and currently there is neither available specific therapeutics to treat nor a vaccine to prevent this disease. Although efforts are being made, development of a vaccine against this disease remains challenging. Hawaii Biotech Inc developed a recombinant subunit envelope protein-based vaccine against all four serotypes produced in Drosophila S2 cells which were transferred over to Merck in 2010. Each subunit of the four dengue serotypes contains the N-terminal 80% of the amino acids comprising the envelope protein (DEN-80E). A Phase 1 study using only monovalent DEN1-80E was done by Hawaii Biotech Inc and most recently, a Phase 1 clinical trial of the tetravalent DEN-80E formulation (V180) was conducted. Here, we report the development of a dose assay for the tetravalent dengue vaccine-containing subunit protein of DEN1-80E, DEN2-80E, DEN3-80E, and DEN4-80E using various separation methods such as HPLC and CE. Based on the results of the comparison, the CZE separation was chosen as the most suitable method to perform the dose assay for the tetravalent dengue vaccine.

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

“…Interestingly, Hamm et al. observed peak tailing of the tetravalent dengue vaccine using different HPMC batches. A binding of the sample molecules to HPMC was assumed.…”

“…Hamm et al. developed a concentration assay for the tetravalent dengue vaccine. Due to the nonavailability of suitable antibody reagents against each dengue serotype, the use of ELISA was difficult.…”

Recent advances in capillary electrophoretic migration techniques for pharmaceutical analysis (2013–2015)

“…As described earlier , CZE has been used for stability indicating methods, e.g. for recombinant human interleukin‐11 and oxytocin , as well as for the dose‐determination of a dengue fewer vaccine candidate .…”

Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals

“…For this purpose, a CZE–CZE–MS approach was published recently . Due to the individual properties of different biopharmaceuticals, various modifications of the initial method from He and colleagues have been published . Goyon et al.…”

“…For this purpose, a CZE-CZE-MS approach was published recently [10]. Due to the individual properties of different biopharmaceuticals, various modifications of the initial method from He and colleagues [8] have been published [11][12][13]. Goyon et al demonstrated an alternative approach for the analysis of charge variants of mAbs using a BGE containing polyethylene oxide for dynamic coating and Bis-tris as a buffering agent.…”