Distinguishing Between Monomeric scFv and Diabody in Solution Using Light and Small Angle X-ray Scattering.

ABSTRACT: Depending on the linker length between the V H and the V L domain, single-chain Fv (scFv) antibody fragments form monomers, dimers (diabodies) or higher oligomers. We aimed at generating a diabody of the anti-MET antibody 3H3 to use it as crystallization chaperone to promote crystallization of the MET ectodomain through the introduction of a pre-formed twofold axis of symmetry. Size exclusion chromatography, however, suggested the protein to be monomeric. Hence, we used scattering techniques applied to solutions to further investigate its oligomerization state. The small angle X-ray scattering (SAXS) curve measured for our protein nicely fits to the scattering curve calculated from the known crystal structure of a diabody. In addition, concentration-dependent photon correlation spectroscop

SUBMITTER: Ludel F 

PROVIDER: S-EPMC6963988 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature