Despite extensive vaccination efforts, certain pathogens continue to pose significant challenges to the poultry industry. Interferons (IFNs) are crucial for innate immune defense against various pathogens. Chicken interferon-alpha (chIFN-α) reportedly has good inhibitory activity against various pathogens. However, its production is often limited by low yields. This study aimed to enhance the production of chIFN-α using an improved baculovirus expression system and evaluate its antiviral efficacy both in vivo and in vitro. A recombinant bacmid carrying two copies of the chIFN-α gene was successfully constructed. The chIFN-α protein was obtained using the modified baculovirus expression system, and exhibited anti-vesicular stomatitis virus (VSV) activity of up to 5.0 × 10