Obtaining and adapting an anti-C3 hybridoma to serum and protein-free medium

Abstract

Coombs serum or polyspecific human antiglobulin reagent allows visualizing the presence of incomplete anti-erythrocyte studies or components of factor C3 of the human complement system adhered to their membranes. It is of great clinical importance to study pathologies such as the disease hemolytic of the newborn, autoimmune hemolytic diseases, among others. Currently, there is a need to cover the hospital demand through national production. The secretory hybridoma of anti-C3 monoclonal studies generated for the production of the Coombs reagent must go through a process of adaptation to serum and protein-free media due to the disadvantages that the traditionally used medium can bring for its cultivation. The current research topic proposes to demonstrate that the selected serum and protein-free media are suitable for hybridoma cultivation. An experimental study was carried out at the Laboratory of Antibodies and Experimental Biomodels in Santiago de Cuba from April to November 2022. Anti-C3 B lymphocytes from the sensitivity of BALB/c mice were obtained. A secreting hybridoma of anti-C3 tests was generated and its adaptation was evacuated in formulations prepared from the MB02, MB03 and MB04 media. Two medium formulations were obtained which, since they are nationally produced, lead to economic advantages. The obtained monoclonal antibodies can be used for the development of a polyspecific human antiglobulin reagent that will satisfy the national hospital demand, guaranteeing the quality of public health care and the substitution of imports.