Expression and bioactivity analysis of TNF30, a TNFα nanobody, in Escherichia coli

Abstract

Antibodies and antagonists targeting tumour necrosis factor alpha (TNFα) have become a number of the best-selling anti-inflammatory drugs. The nanobody is a single domain antibody derived from the variable domain of a heavy-chain antibody. However, so far, there are still no reports on the recombinant expression of TNFα nanobodies. Here, TNF30, the key anti-TNFα nanobody domain of ozoralizumab, was cloned, expressed in Escherichia coli, and then purified with the Ni2+-chelating affinity chromatography. The western blot results showed that the recombinant TNF30 has strong binding capability with TNFα. Furtermore, the pharmacological and histopathological analysis in the carrageenan-induced mice paw oedema demonstrated that the recombinant nanobodies had potential anti-inflammatory activity in vivo. These works provided a novel approach for the production of TNF30 in E. coli, and might establish a foundation for the industrial production of the drug and the development of more active derivatives against TNFα in the future.

Keywords

• Tumour necrosis factor
• nanobody
• recombinant expression
• arthritis