Provided by the present invention are an antibody against human TIM-3 or an antigen-binding fragment thereof, and further provided are a nucleic acid molecule encoding the antibody, an expression vector for expressing the antibody and a host cell, and a method for producing the antibody. Further provided by the present invention are a pharmaceutical composition comprising the antibody or an antigen-binding fragment of the antibody, and an application of the pharmaceutical composition in the preparation of a medicine, the medicine is used for preventing and/or treating various diseases (including tumors, infectious diseases and autoimmune diseases).