Expression of viral vectors or other therapeutic proteins in HEK 293 cells is a widely used procedure for obtaining sufficient quantities of a desired virus or protein. The manufacturing and purification process of these products leaves the potential for impurities by host cell proteins (HCPs) from HEK 293 cells. HCP impurities can result in adverse toxic or immunological reactions and thus it is desirable to reduce these impurities to the lowest levels practical.
The polyclonal antibodies used in this kit have been generated against and affinity-purified using a mild lysate of HEK 293 cells to obtain HCPs typically encountered in initial product recovery steps. This kit can be used as a process development tool to monitor the optimal removal of host cell impurities as well as in routine final product release testing when comprehensively characterized using samples from your process. The orthogonal methods of Antibody Affinity Extraction (AAE) and western blot were used to determine reactivity to individual HCPs in a mild lysate of HEK 293. AAE indicated reactivity to ~70% of the individual 2-D PAGE fractionated spots presumed to be HCP. In terms of the total mass of HCP, the anti-HEK 293 antibody reacts to those HCPs that represent >90% of the total HCP mass in the lysate sample. This ELISA Kit 'F650R' replaces the previous F650 kit while incorporating some improvements in performance. The F650R kit uses a new antibody with comparable reactivity to the previous F650 antibody and thus you can expect similar HCP levels between the two kits. The kit has also been successfully used to identify HCPs from HEK 293T cells.