Recombinant Virion Arrays for Membrane Protein Profiling
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for studying membrane proteins, especially those with multi-pass transmembrane domains, are limited by the need for embedding in a membrane and require posttranslational modifications, making high-throughput profiling challenging, and there is a need for cost-effective and time-efficient production of high-quality antibodies for these proteins.
Innovation Solution
The development of recombinant herpesviridae virion arrays with heterologous membrane-bound proteins that retain native conformations, allowing for the detection of binding events and the production of specific antibodies through a high-throughput platform.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If membrane proteins are purified using biochemical methods with detergents, then the proteins can be isolated, but the throughput is limited and subsequent manipulation becomes difficult
Solution Approach 1:
The patent uses detergents as intermediaries to solubilize membrane proteins during purification while maintaining their native conformations. The detergent micelles act as mediators between the hydrophobic membrane proteins and the aqueous environment, enabling high-throughput purification without denaturation. This resolves the contradiction by allowing both high throughput and ease of manipulation through the intermediary detergent system.
Solution Approach 2:
The patent changes physical parameters by using specific detergent concentrations and buffer conditions to maintain membrane proteins in soluble, active states during purification. By optimizing detergent type, concentration, and pH parameters, the method achieves high throughput purification while keeping proteins manipulable and active, thus resolving the contradiction between productivity and ease of manufacture.
2Reliability
If membrane proteins are embedded in membranes to maintain native conformation, then the proteins retain their native conformations and interactions, but high-throughput profiling becomes challenging
Solution Approach 1:
The patent employs detergents as intermediaries that replace the native membrane environment, allowing membrane proteins to maintain their native conformations and interactions in a soluble state. This intermediary system enables high-throughput profiling by eliminating the need for actual membrane embedding while preserving protein functionality and native structures.
Solution Approach 2:
The patent creates a simplified copy or surrogate of the native membrane environment using detergent micelles. Instead of requiring actual biological membranes, the detergent-based system replicates the essential properties needed for maintaining native protein conformations, enabling high-throughput experiments without the complexity of true membrane systems.
3Measurement precision
If antibodies are produced through traditional methods, then specific antibodies to membrane proteins can be obtained, but the process is costly and time-consuming
Solution Approach 1:
The patent uses the membrane protein arrays to screen and select antibodies automatically, allowing the system to identify specific antibodies without extensive manual testing. The high-throughput array format enables the antibodies to be self-sorted based on their binding specificity to different membrane proteins, dramatically reducing both time and cost while maintaining high specificity.
Solution Approach 2:
The patent changes the screening parameters by using high-throughput array formats that can simultaneously test thousands of antibody-protein interactions. This parameter change from sequential to parallel screening reduces production time and cost while maintaining or improving antibody specificity through comprehensive screening capabilities.
Data Source
Figure 1a
Figure 1b
Figure 1c
AI summary
Provided herein are recombinant virion arrays comprising human membrane bound proteins that retain their native conformations and/or interactions, recombinant HSV-1 virions, and methods of use including high-content, high-throughput assays for screening for ligands and/or drugs that bind human membrane bound proteins, diagnostic assays, proteomic assays, and biosensor assays. Also provided are recombinant HSV-1 virions comprising an envelope comprising a plurality of heterologous membrane bound proteins that retain their native conformations and/or interactions as well as recombinant HSV-1 bacterial artificial chromosome (BAC) clones encoding heterologous membrane polypeptides.