The present invention relates to the field of biological products, and in particular relates to a vaccine freeze-drying protective agent containing no gelatin and human albumin. The vaccine freeze-drying protective agent comprises 1.5%-10% of sucrose, 1.5%-3.5% of dextran, 1%-2% of sorbitol, 0.8%-1.2% of sodium glutamate and 0.2% to 1% of L-arginine in a vaccine semi-finished product, a matrix liquid for preparation of the vaccine freeze-drying protective agent is 199 culture medium or PBS buffer solution or water for injection, the vaccine freeze-drying protective agent contains no gelatin and human albumin, and the vaccine semi-finished product is a liquid vaccine before freeze-drying. The present invention also provides the use of the vaccine freeze-drying protective agent. When the vaccine freeze-drying protective agent is used for the preparation of a freeze-dried vaccine, the vaccine freeze-drying protective agent can improve the vaccine stability during freeze-drying and storage processes, greatly improves the freeze-dried vaccine safety for human, and reduces adverse effects of the vaccine.