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This invention discloses a method for preparing chlorantraniliprole nanobodies, belonging to the field of biorecognition material preparation for pesticide residue detection. The method involves screening chlorantraniliprole-specific nanobodies using bioinformatics analysis, cloning them into the pET-22b(+) expression vector, transforming them into *E. coli* BL21(DE3) competent cells, and achieving soluble expression of the nanobodies via IPTG induction. High-purity chlorantraniliprole nanobodies are then obtained through periplasmic purification combined with nickel column affinity chromatography. This invention eliminates the need for animal immunization experiments such as alpaca screening through bioinformatics, significantly shortening the nanobodies preparation cycle and reducing costs. The use of the pET-22b(+) prokaryotic expression system combined with an 18℃ low-temperature induction strategy achieves soluble expression of chlorantraniliprole nanobodies. After induction, the target protein band can be clearly detected by SDS-PAGE electrophoresis (see Figure 1), improving the correct folding ratio of the antibody.