The invention provides a preparation method of anti-human lymphocyte immunoglobulin, which comprises the steps of: immunizing healthy pigs with human peripheral blood T lymphocytes or human thymocytes, collecting and separating pig plasma, making it with 20% ethanol, and using A low-temperature high-speed centrifuge or a filter press separates the precipitate of component I+II+III, and after making it with 14% ethanol, separates and collects the supernatant, and then makes it with 25% ethanol, separates the precipitate of component II, and then The anti-human T cell porcine immunoglobulin can be obtained through steps such as immunoglobulin purification, configuration and virus inactivation. Compared with the currently used ammonium sulfate salting-out method, the process of the present invention has the advantages of mass production, simple operation and easy mastery, short production cycle, less reagents used, low cost, no environmental pollution, etc., and overcomes the difficulties existing in the existing process. Large-scale production, low output, large waste, environmental pollution, high production cost, complicated production process, various chemical reagents used, long production cycle, high equipment occupancy rate, etc.