Journal: Vaccines

Article Title: Establishment and Validation of a Cell-Based Relative Potency Method for Respiratory Syncytial Virus mRNA Vaccine Drug Substance

doi: 10.3390/vaccines14050401

Figure Lengend Snippet: Optimization of cell-related conditions for the RSV mRNA potency assay. ( A ) Dose–response curves of RSV mRNA transfection in different cell lines (293T, Vero, HEp-2, HepG2, and A549). Cells were transfected with increasing amounts of RSV mRNA (0.0002–0.48 µg/well), and RSV prefusion F protein expression was measured by ELISA as OD 450 –OD 630 . The corresponding EC 50 and slope values are summarized in the inset table. ( B ) Effect of HepG2 cell seeding density on RSV prefusion F protein expression. HepG2 cells were seeded at 0.5 × 10 5 , 1.0 × 10 5 , or 2.0 × 10 5 cells/well and transfected with increasing amounts of RSV mRNA (0.0002–0.48 µg/well). Protein expression was measured by ELISA as OD 450 –OD 630 .

Article Snippet: Cell lines used for mRNA transfection included the human liver carcinoma cell line HepG2 (American Type Culture Collection, ATCC, Manassas, VA, USA), human non-small cell lung carcinoma cell line A549 (ATCC), African green monkey kidney epithelial cell line Vero (ATCC), human epidermoid carcinoma type 2 cell line Hep-2 (ATCC), and human embryonic kidney cell line 293T (China Center for Type Culture Collection, CCTCC, Wuhan, China).

Techniques: Potency Assay, Transfection, Expressing, Enzyme-linked Immunosorbent Assay