473766 Development of Thermostable Affinity Reagents for Low-Cost, Point-of-Care Diagnostics
We here report the development of thermostable affinity reagents based on a DNA-binding protein isolated from the hyperthermophilic bacteria S. solfataricus. This scaffold was used to develop a model binding protein, which has been assessed for its surface-bound activity, its utility within the dilute antigen regime, and its activity retention under thermal challenge. The demonstrated kinetic stability of this species is nearly three orders of magnitude greater than that of standard polyclonal antibodies, and the species can be facilely produced in high molar yields via bacterial expression and purified in a single chromatographic step. Using yeast surface display and flow cytometric analysis, rationally-designed mutagenesis libraries have also been screened for potential binders to four TB biomarkers identified in the urine of infected patients via mass spectrometry. These studies have confirmed the scaffold’s potential applicability to a broad suite of disease biomarkers, and have validated the suitability of this scaffold as a replacement for antibodies in in vitro point-of-care diagnostic devices.
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