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Antibodies are glycoproteins, which consist of two types of domains light and heavy chains, each of which is composed of variable and constant domains. Rituximab is a monoclonal antibody used in the treatment of lymphomas and its mechanism of action is based on the interaction of CD20 membrane protein, expressed on B lymphocytes, guiding them to depletion. Single-chain fragments variable (scFv) are composed by the variable domains of antibody heavy and light chains, linked by a flexible polypeptide, making them easier to manipulate on protein engineering techniques, such as rational design, where amino acids are chosen by in silico structural and energetic analysis and changed by site-directed mutagenesis in order to set a better or new biological function.

Construction, by rational design, and initial characterization of affinity mutants of Rituximab fragment antibody