title: Methodology for preservation of yeast-bound single chain fragment variable antibody affinity reagents.
abstract: Readily accessible affinity reagents are critical to the validation of biomarkers and to the development of new diagnostic tests. As alternatives to monoclonal antibodies, yeast-bound single chain fragment variable antibody (yeast-scFv) can be rapidly selected from yeast display libraries. An important characteristic for any diagnostic reagent is its stability or ability to store it. A lyophilization procedure that has extended the shelf life of yeast-scFv by a factor of ≥10-fold relative to previous reports is reported. Real time stability for three yeast-scFv clones to three distinct Entamoeba histolytica potential diagnostic antigen targets for one year at room temperature as well as at 37°C and 45°C. Retention of full binding activity and specificity of the yeast-scFv clones for their cognate antigens is shown by flow cytometry. Lyophilization can easily be carried out in batches and in single-use vials.
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: Maria
authors.initials: M
authors.lastname: Kahn
authors.affiliation: Eurofins Lancaster Labs, Thousand Oaks, CA, United States.
authors.forename: Scott
authors.initials: S
authors.lastname: Priddy
authors.affiliation: PATH, Seattle, WA, United States.
authors.forename: Marcus
authors.initials: M
authors.lastname: Estrada
authors.affiliation: University of Washington, Seattle, WA, United States.
authors.forename: Lauren
authors.initials: L
authors.lastname: Spadafora
authors.affiliation: University of Washington, Seattle, WA, United States.
authors.forename: Gerard A
authors.initials: GA
authors.lastname: Cangelosi
authors.affiliation: PATH, Seattle, WA, United States. Electronic address: gdomingo@path.org.
authors.forename: Gonzalo J
authors.initials: GJ
authors.lastname: Domingo
mesh_terms: D004355:Drug Stability; D004356:Drug Storage / Q000379:methods*; D005434:Flow Cytometry; D005612:Freeze Drying; D011933:Reagent Kits, Diagnostic; D057127:Single-Chain Antibodies; D015003:Yeasts
chemical_list: D011933:Reagent Kits, Diagnostic; D057127:Single-Chain Antibodies
keywords: Diagnostics; Neglected tropical diseases; Yeast display