GLUT4 Antibodies via VLP Immunization for Conformational Detection

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Solution Overview

Problem

Current methods lack effective tools to study and detect the conformational states of GLUT4 glucose transporters on the cell surface, particularly due to the scarcity of tools for measuring wild-type GLUT4 and the difficulty in maintaining the native structure of multispanning membrane proteins for antibody production.

Innovation Solution

Development of antibodies that specifically bind to the native conformational states of GLUT4, including inward-open and outward-open states, generated using virus-like particles and a divergent host species like chickens, which recognize conformational epitopes and inhibit GLUT4 function.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional antibodies recognizing linear epitopes are used, then Western blotting applications are effective, but flow cytometry and surface translocation measurement are ineffective

Engineering Contradiction:

ImproveGLUT4 surface translocation detectionVSAvoidantibody production difficulty

Core Design Contradiction:

Measurement precisionVSEase of manufacture

Solution Approach 1:

The patent applies preliminary action by pre-conforming the GLUT4 antigen into its native conformational state within virus-like particles before antibody exposure. This allows the immune system to generate antibodies that recognize the native conformational epitopes directly, resolving the contradiction between measurement precision for surface translocation and the difficulty of maintaining native structure during antibody production

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent uses virus-like particles as an intermediary carrier to present the GLUT4 antigen in its native conformational state. This intermediary structure maintains the three-dimensional folding and conformational integrity of GLUT4 during immunization, enabling the production of antibodies that specifically recognize surface-exposed conformational epitopes rather than linear sequences

Inventive Principle:

Principle #24Intermediary (Mediator)

2Reliability

If the full length GLUT4 protein is used in membrane context for immunization, then native conformational antibodies can be generated, but the small extracellular loops form a small target antigen reducing immune reactivity

Engineering Contradiction:

Improvenative conformation recognitionVSAvoidantibody yield

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent merges the GLUT4 antigen with virus-like particle structures to create a hybrid immunogen. This combining approach amplifies the immunogenicity by presenting multiple copies of the conformational epitopes on the VLP surface, while maintaining the native structure recognition requirement. The VLP acts as a scaffold that concentrates and displays the small extracellular loop epitopes in a highly immunogenic format

Inventive Principle:

Principle #5Merging (Combining)

3Ease of manufacture

If traditional animal hosts are used for immunization, then standard antibody production procedures can be followed, but GLUT4 high conservation among mammals makes these hosts poor candidates

Engineering Contradiction:

Improveantibody production procedureVSAvoidconformational epitope specificity

Core Design Contradiction:

Ease of manufactureVSMeasurement precision

Solution Approach 1:

The patent applies inversion by using a distantly related species (chicken) instead of a closely related mammalian host for immunization. This reverse approach exploits the lower sequence homology between chicken and human GLUT4 to generate antibodies with higher conformational epitope specificity, overcoming the limitation of traditional mammalian hosts where high conservation reduces antibody discrimination capability

Inventive Principle:

Principle #13The other way round (Inversion)

Data Source

PatentUS11834497B2Glucose transporter 4 antibodies, methods of making the same, and uses thereof

Publication Date: 2023.12.05 INTEGRAL MOLECULAR INC

AI summary

Antibodies and compositions against GLUT4 and uses thereof are provided herein.