Optimal Fusion of Antibody Binding Domains Resulted in Higher Affinity and Wider Specificity

Dong J, et al., pii: S1389-1723(15)00141-3, J Biosci Bioeng, 2015

With the aim of improving the affinity and specificity of binding domains, the authors generated novel fusion proteins (PAxPG) with a flexible linker between the two immunoglobulin binding domains derived from Protein A and Protein G. Once constructed, these fusion proteins were evaluated for their binding affinity to human Fab and certain mouse IgG subclasses (IgG1). A Pall ForteBio BLItz system equipped with Streptavidin biosensors was used in the analyses of binding interactions. Biotinylated PAxPG were captured onto Streptavidin biosensors and screened against the analytes such as the Human anti-vimentin Fab, human IgG1 Fc, and mouse IgG1. The data offers crucial insights into the linker lengths that can be used in the fusion proteins.

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