Biotinylation Eliminates the Intermediate State of Top7 Designed with an HIV-1 Epitope

Broadly neutralizing antibodies against HIV-1 are rare with the 2F5 antibody being one of the most protective. Insertion of an antibody epitope into a stable and small protein scaffold overcomes many of the obstacles found to produce antibodies. However, the design leads to grafting of epitopes that may cause protein aggregation. Here, I investigated the 2F5 epitope grafted into the Top7 as the scaffold in which the resulting immunoreactive protein precipitates along the storage time, as opposed to its completely soluble biotinylated version. Molecular dynamics showed that biotinylation eliminates the intermediate state of the scaffold-epitope Top7-2F5 by switching a noncooperative to a cooperative folding. The aggregation propensity of the Top7-designed proteins is examined in light of thermodynamic cooperativity and kinetic traps along the decreasing depth of the intermediate ensemble in the free energy landscape. This protocol may predict stable and soluble scaffold-epitopes with the purpose of composing novel therapeutic and diagnostic platforms.

Automated summary

This study investigated the grafting of the HIV-1 broadly neutralizing antibody epitope 2F5 into the Top7 protein scaffold. It was found that the grafted protein precipitates during storage, unlike its biotinylated version which remains soluble. Molecular dynamics revealed that biotinylation eliminates the intermediate state and induces a cooperative folding. The findings contribute to the prediction of stable and soluble scaffold-epitopes for the development of novel therapeutic and diagnostic platforms.