Anal Biochem. 2018 Jan 12. pii: S0003-2697(18)30007-1. doi: 10.1016/j.ab.2018.01.006. [Epub ahead of print]
Quantification of a recombinant antigen in an immuno-stimulatory whole yeast cell-based therapeutic vaccine.
Wang J1, Stenzel D1, Liu A1, Liu D1, Brown D1, Ambrogelly A2.
Author information
1
Biologics Analytical Operations, Pharmaceutical & Biologics Development, Gilead Sciences, 4010 Ocean Ranch Blvd, Oceanside, CA, 92056, United States.
2
Biologics Analytical Operations, Pharmaceutical & Biologics Development, Gilead Sciences, 4010 Ocean Ranch Blvd, Oceanside, CA, 92056, United States. Electronic address: [email protected].
Abstract
Therapeutic vaccines represent an emerging class of immune-modulatory treatments for cancer, infections, and chronic diseases. One such vaccine was designed as an immune stimulator of the T cell response against HBV antigens to eliminate HBV infected cells and offer a therapeutic avenue to treat patients suffering from chronic hepatitis B infection. Whole deactivated Saccharomyces cerevisiae cells expressing a recombinant fusion of HBV X, S and Core antigens elicit T cell responses in mice and activate human T cells linked with viral clearance. As the therapeutic efficacy of the yeast-based vaccine relies on the production of the recombinant antigen, analytical methods designed to accurately and precisely quantitate the fusion protein in the midst of all the yeast proteins are necessary. We report the development and characterization of western blot, quantitative ELISA and mass spectrometry based orthogonal methods to support the assessment of manufacturing consistency.
KEYWORDS:
Antigen; Core antigen; ELISA; HBV; Immuno modulatory; MRM; S antigen; Saccharomyces cerevisiae; Tarmogen(®); Whole cell therapeutic vaccine; X antigen
