Thermonobodies: A Novel Thermostable FN3 Binding Protein Scaffold

SUMMARY

• Monobodies are synthetic binding proteins based off a human fibronectin type 3 (FN3) scaffold that have demonstrated their utility in applications such as the modification of enzyme specificity and therapeutics for topologically challenging targets. Thermostability limits shelf life, oral bioavailability in therapeutic applications, and high temperature industrial reaction engineering applications. 
• “TherMonobodies” are Monobodies engineered from a hyperthermophillic archaeon FN3 scaffold with further structural modifications to boost thermostability over the original human FN3 scaffold
• TherMonobody libraries have been engineered with: 1) a loop-based diversity for recognition of cleft-like epitopes protruding from the target and 2) a side-and-loop based diversity for recognition of a more continuous surface on the target. The scaffold is engineered for favorable expression in both the yeast and phage display formats.
• In a proof-of-concept study both loop and side-and-loop TherMonobody libraries were screened against ySUMO, MBP, GFP, Abl SH2, SHP2 N-, and C-SH2 model targets. The resulting clones had Tms ranging from 50°C-86°C, a 13°C increase over the original Monobody scaffold.

FIGURE

ADVANTAGES

ADVANTAGES

• Increased thermostability over traditional Monobodies
• Ten-fold smaller than traditional antibodies
• Alternate binding mechanism from other immunoglobins is amenable to topologically challenging targets

APPLICATIONS

• Industrial reaction engineering and catalysis
• Diagnostics
• Research tools

PUBLICATIONS

• TherMonobides, synthetic binding proteins based on a hyperthermophillic fibronectin type III domain. Tanka, et al. In preparation.

• PCTUS2019/018866.
• For more information on enzyme modification with Monobodies, see also: Novel Platform for Allosterically Modifying Enzyme Specificity (14-T-118)