Production of anti-L.tropica Nanobodies fused with GFP protein and assessment their sensitivity

Abstract

Nanobodies derived from the heavy chain antibodies of the family of camelidea are one of the most important recombinant antibodies due to their special structural properties, as well as the possibility of cloning them into different features and forms. Since fusing antibodies with green fluorescent protein GFP is a way to be used in diagnostic or therapeutic applications, this work aimed to fuse anti L. tropica nanobodies which diffuse in Syria with GFP protein, optimize expression conditions, and study the advantages of this fusion. The gen of anti L. tropica nanobodies were cloned within PMES4-GFP, and when testing different conditions for the expression of the fusing nanobodies, their accumulation was shown in inclusion bodies, temperature 22 ° C and 0.5 mM concentration of IPTG were the optimal conditions for expression. Significantly, the sensitivity of the conjugated nanobodies and their ability to identify low concentrations of leishmaniasis antigens 30 ng/ml compared with non- conjugated nanobodies except for NbLTC24 showed no difference whether they were fused with GFP or not.