Hsieh, Ching-LinRush, Scott APalomo-Sanz, ConcepcionChou, Chia-WeiPickens, WhitneyMas-Lloret, VicenteMcLellan, Jason S2022-05-242022-05-242022-03-14Nat Commun. 2022 Mar 14;13(1):1299.http://hdl.handle.net/20.500.12105/14467The human metapneumovirus (hMPV) fusion (F) protein is essential for viral entry and is a key target of neutralizing antibodies and vaccine development. The prefusion conformation is thought to be the optimal vaccine antigen, but previously described prefusion F proteins expressed poorly and were not well stabilized. Here, we use structures of hMPV F to guide the design of 42 variants containing stabilizing substitutions. Through combinatorial addition of disulfide bonds, cavity-filling substitutions, and improved electrostatic interactions, we describe a prefusion-stabilized F protein (DS-CavEs2) that expresses at 15 mg/L and has a melting temperature of 71.9 °C. Crystal structures of two prefusion-stabilized hMPV F variants reveal that antigenic surfaces are largely unperturbed. Importantly, immunization of mice with DS-CavEs2 elicits significantly higher neutralizing antibody titers against hMPV A1 and B1 viruses than postfusion F. The improved properties of DS-CavEs2 will advance the development of hMPV vaccines and the isolation of therapeutic antibodies.engVoRhttp://creativecommons.org/licenses/by/4.0/Protein vaccinesVirologyX-ray crystallographyAtribución 4.0 Internacional35288548131129910.1038/s41467-022-28931-32041-1723Nature Communicationsopen access