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This invention discloses a method for efficient soluble expression of classical swine fever virus (CSFV) E2 protein and a method for preparing highly active monoclonal antibodies, belonging to the fields of animal disease detection and bioengineering technology. This invention, through precise optimization of dual sequences, targeted modification of molecular chaperone vectors, synergistic dual molecular chaperone combinations, temporal induction processes, and a dual screening system, balances antigen activity and translation efficiency, improves induction response and expression efficiency, and precisely matches the folding sequence to obtain E2 protein with a soluble content of approximately 86% and a purity ≥95%. The 3B7 monoclonal antibody prepared using this protein as an immunogen showed the best performance, with an ascites titer of 1:96000, and neutralizing titers of 1:5120 and 1:4096 against CSFV Shimen strain and prevalent subtypes, respectively. The titer decreased by only 4.2% after 3 months of refrigeration at 4°C, and it specifically recognizes CSFV types 1 and 2 without cross-reactivity with BVDV / BDV. This invention solves the pain points of existing technologies, has low industrialization costs, and provides important technical support for the precise prevention and control of classical swine fever.