In the field of cryobiology, there are a large number of publications devoted to the study of the most labile lipid composition of gametes and their membranes. However, less attention is paid to the state of the protein component of cells. Therefore, the purpose of the presented paper was to study the protein spectrum and its changes in the process of cryopreservation of bull and rooster sperm. The presence of albumins, alpha, beta and gamma globulins was determined by the spectrophotometric method. Most of the proteins are contained in the seminal plasma, in gametes the total protein content is slightly lower. Of the seminal plasma proteins, most contain albumins, and of the gamete proteins, most contain gamma globulins (in the sperm of both animal species). With the passage of technological stages, the number of some proteins increases, while of others it decreases. This indicates about the destruction of protein structures, violation of the permeability of biological membranes and changes in the chromatographic mobility of proteins. From the data obtained, it was found that the protein spectrum of bull and rooster sperm is very different. If alpha globulins (18.2±1.84 mg/billion) prevail in the plasma of the bull`s sperm, and gamma globulins (16.8±3.27 mg/billion) prevail in the gametes, then in rooster sperm, both in plasma and gametes, most of all is albumin (6.0±2.45 and 11.8±0.47 mg/billion, respectively).