The invention provides a method for adopting a means of zooblast fed-batch culture to highly efficiently express recombinant proteins. The method of the invention selects the means of zooblast fed-batch culture to express the recombinant proteins, and controls cell culture temperature and PH value of different ranges in different phases of fed-batch culture so as to reach the aim of highly efficient expression of the recombinant proteins. The invention has the advantages of simple and convenient operation, easy implementation, and improvement of protein output simultaneously without improvingthe cost. The invention is suitable for large-scale zooblast culture and particularly suitable for being applied in producing various bioactive substances like monoclonal antibodies, fused proteins and so on and various gene recombinant proteins.