Towards a Universal Disulphide Stabilised Single Chain Fv Format: Importance of Interchain Disulphide Bond Location and vL-vH Orientation

Weatherill, E.E., et al., 25(7), 321-329, Protein Engineering, Design and Selection, 2012

Five disulphide bond locations were introduced into one scFv in order to compare their relative effects on expression, thermal stability, % monomer formation and retention of antigen binding. The Octet system was used to determine the expression levels of the scFv with different bond modifications using a Streptavidin Biosensor coupled to biotinylated Protein L to quantitate the scFv forms. The expression levels of scFvs were not significantly affected in various disulphide bond locations with the exception of vH101-vL46 bond location.

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