Initial development of biosimilar immune checkpoint blockers using HEK293 cells

Publication date: June 2020Source: Protein Expression and Purification, Volume 170Author(s): Michael Bernardes Ramos, Anna Erika Vieira de Araújo, Cristiane Pinheiro Pestana, Ana Paula Dinis Ano Bom, Renata Chagas Bastos, Aline de Almeida Oliveira, Patrícia Cristina da Costa Neves, Haroldo Cid da Silva JuniorAbstractAntibodies that block interaction of immune checkpoint receptors with its ligands have revolutionized the treatment of several cancers. Despite the success of this approach, the high cost has been restricted the use of this class of drugs. In this context, the development of biosimilar can be an important strategy for reducing prices and expanding access after patent has been dropped. Here, we evaluated the use of HEK293 cells for transient expression of an immune checkpoint-blocking antibody as a first step for biosimilar development. Antibody light and heavy chain genes were cloned into pCI-neo vector and transiently expressed in HEK293 cells. The culture supernatant was then subjected to protein A affinity chromatography, which allowed to obtain the antibody with high homogeneity. For physicochemical comparability, biosimilar antibody and reference drug were analyzed by SDS-PAGE, isoelectric focusing, circular dichroism and fluorescence spectroscopy. The results indicated that the both antibodies have a high degree of structural similarity. Lastly, the biosimilar antibody binding capacity to target receptor was shown to be similar to reference product in EL...
Source: Protein Expression and Purification - Category: Biochemistry Source Type: research