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US 11,892,457 B2Granted
Proteoliposome-based ZnT8 self-antigen for type 1 diabetes diagnosis
AssigneesThe Johns Hopkins University, The Board of Trustees of the Leland Stanford Junior University
InventorsDax Fu, Chengfeng Merriman, Hongjie Dai, Hao Wan
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Abstract
Methods of detecting ZNT8 antibodies in serum are described. The methods include proteoliposomes comprising a transmembrane domain (TMD) and a cytosolic domain (CTD) of ZnT8 proteins exposed on the exterior of the proteoliposome; serum comprising antibodies targeting the ZnT8 proteins; and labelled captured autoantibodies that bind to ZnT8 antibodies.
Claims16
A method of detecting ZnT8 antibodies in serum comprising:
- (a) providing a collection of proteoliposomes having an interior and an exterior wherein some proteoliposomes have protein sequences of a transmembrane domain (TMD) of a zinc transporter 8 (ZnT8) on its exterior and other proteoliposomes comprise protein sequences of a cytosolic domain (CTD) of a ZnT8 protein exposed on its exterior,
- wherein the protein sequences of a TMD and a CTD are part of a full length ZnT8 protein,
- wherein the full length ZnT8 protein is a 325R polymorphic variant of human ZnT8,
- wherein the collection of proteoliposomes is prepared by dehydrating soluble ZnT8-containing proteoliposomes followed by rehydration of the dehydrated ZnT8-containing proteoliposomes, and
- wherein the collection of proteoliposomes is bound to a plate or chip;
- (b) adding serum comprising antibodies targeting the protein sequences of TMD and/or CTD forming proteoliposomes bound to the antibodies;
- (c) adding a labelled secondary antibody that binds to the antibodies bound to the proteoliposomes;
- and
- (d) measuring the amount of antibodies targeting the protein sequences of TMD and/or CTD bound to the proteoliposomes.
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Dependencies are inferred from the claim text.