Rotavirus is the most common cause of diarrhea in infants and young children under five years old. The VP6 protein of rotavirus was considered as an important antigen in the development of novel vaccine generation and diagnostic tools. Polyclonal antibodies of VP6 antigen could be used to generate a rapid test based on the lateral flow immunoassay. In the present study, we showed the results of synthesis of polyclonal antibodies against recombinant VP6 protein of rotavirus. Recombinant VP6 protein was strongly produced by E. coli in suitable conditions. The properties of recombinant VP6 protein were characterized by Western blot and ELISA methods. Pure recombinant VP6 protein was used as an antigen to inject into the rabbits. The recombinant VP6 protein was highly immunogenic in rabbits. Anti-VP6 antibodies were purified from the rabbit serum using the Protein A affinity chromatography. The specificity of the antibody towards its antigen was characterized by Western blot and ELISA immunoassay. The purity of antibody was reached as 97 %. The efficiency of antibody production in rabbit was 3.94 mg/ml serum. The amount and quality of the obtained anti-rotavirus VP6 antibody was suitable for application in the development of the diagnostic kits.