Investigation of the Effect of Arginine and Glutathione on Recovery of a Single Domain Antibody Produced in Bacteria in Inclusion Bodies

Single domain antibodies (nanobodies) are the antigen binding domain of heavy chain only antibodies derived from camelids and sharks. Standard antibodies have heavy and light chains, and the antigen binding region is formed by the combination of variable heavy and variable light chain domains. Single domain antibodies are as good binders to their respective antigens as the combination of variable heavy and light chains. Due to their chemical and thermal stability, small size, and economic benefits, there is increasing interest in nanobodies for research use and from industry. In this article, we investigated the effect of arginine and a mixture of oxidized and reduced glutathione for recovering a nanobody produced in inclusion bodies in E. coli. Nanobody protein is solubilized in 6M urea buffer from the cell lysate. After a Ni-NTA chromatography, nanobody containing fractions were first diluted in different concentrations of arginine and/or reduced and oxidized glutathione containing buffers, followed by dialysis against a buffer to fold the protein. The best recovery yield was obtained in the presence of 400mM arginine. Nanobodies are important molecules in biotechnology and medicine, and, this study investigated ways to improve their production yield.