The invention discloses a highly functional and active yellow fever virus human monoclonal antibody and an application thereof, belonging to the technical field of medicine. In the present invention, the yellow fever virus E protein expressed by Escherichia coli is used as the antigen, and memory B cells that can specifically bind to the yellow fever virus E protein are screened from the PBMCs of a convalescent patient through flow sorting, and then the single B cells screened are screened. The cells are subjected to RT-PCR and PCR amplification to obtain variable region fragments of the antibody, which are further linked with the constant region into an expression vector. After being expressed and purified in mammalian cells, a series of functional tests were carried out to obtain a human monoclonal antibody capable of protecting against yellow fever virus infection. The antibody and antigen affinity is 0.588nM, has strong neutralizing yellow fever virus activity, IC50 is 3.7ng / ml, can completely protect mice from lethal dose of yellow fever virus attack, with clinical treatment and prevention of yellow fever virus Application value of heat virus.