Disclosed is a second-order culture method for producing a suspended Vero cell vaccine, wherein same comprises the following steps: suspension culture growth of Vero cells, inoculation of a virus, dilution or addition of a production medium, secondary growth of cells, and optionally harvesting and purifying the virus fluid to produce the vaccine.

Disclosed is a second-order culture method for producing a suspended Vero cell vaccine, wherein same comprises the following steps: suspension culture growth of Vero cells, inoculation of a virus, dilution or addition of a production medium, secondary growth of cells, and optionally harvesting and purifying the virus fluid to produce the vaccine.

A detection of optimal genetic recombinants used to prepare target proteins, with assessment of their target-specific upstream productivity, genetic stability and means to optimize target protein downstream purification using customizable fluorescent tags. A scarless removable protein fusion makes it possible to identify recombinants of Pichia pastoris with optimal performance in heterologous protein production.