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1. (WO2018039802) ALLOSTERIC SPLIT TREHALASE BIOSENSOR

Pub. No.:    WO/2018/039802    International Application No.:    PCT/CA2017/051033
Publication Date: Fri Mar 09 00:59:59 CET 2018 International Filing Date: Sat Sep 02 01:59:59 CEST 2017
IPC: C12Q 1/00
C12M 1/34
C12Q 1/26
C12Q 1/34
G01N 21/75
G01N 27/403
G01N 33/53
Applicants: CREATIVE PROTEIN SOLUTIONS INC
Inventors: DE BUCK, Jeroen
Title: ALLOSTERIC SPLIT TREHALASE BIOSENSOR
Abstract:
The present disclosure relates to a method referred to herein as the "split trehalase assay biosensor" (also referred to herein as "STIGA") is based on the use of engineered E.coli trehalase to detect analytes such as antibodies in a sample. The trehalase is engineered in a way such that the enzyme is split into two inactive fragments (N-terminal fragment H and C-terminal fragment A) with antigens fused to both fragments. When bivalent antibodies react specifically with the fused antigens, two inactive trehalase fragments are brought in close proximity to restore the activity of trehalase. The restored trehalase will hydrolyze trehalose into two glucose molecules which can be measured using existing glucose detection methods such as glucometer, Benedict's reagent, or ACCU- CHEK AVIVA® glucose test strips.