Phage Display Library for High-Throughput Host-Microbe Interaction Screening
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Solution Overview
Problem
Current methods for identifying host proteins involved in microbial interactions are limited by low throughput and inability to detect secreted proteins that mediate complex interactions, such as neutralization or exclusion from the mucus layer, necessitating a high-throughput assay to profile host-microbe interactions effectively.
Innovation Solution
A method using a display library of yeast cells expressing various proteins or peptides, such as nanobodies, to interact with microbial species, followed by selection and identification of bound molecules, enabling the detection of host proteins that interact with microbes and potential therapeutic targets.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If split-domain proteins or other extracellular interaction methods are used, then sensitivity for protein-protein interactions is improved, but device complexity and ease of operation deteriorate due to engineering requirements and heterologous expression systems
Solution Approach 1:
The patent uses phage display technology to create a copy of the host extracellular proteome on phage particle surfaces. Instead of engineering complex fusion proteins or heterologous expression systems, the invention displays purified host proteins on phage particles, simplifying the system while maintaining interaction detection capability. This copying approach allows direct screening of protein-microbe interactions without complex engineering.
2Ease of operation
If pairwise interaction methods are used, then ease of operation is improved, but productivity deteriorates due to inability to detect multi-subunit complexes and require prediction, cloning, and expression
Solution Approach 1:
The patent creates a universal library of phage particles displaying multiple host extracellular proteins simultaneously. This single system can detect multiple types of interactions including protein-protein, protein-peptide, and protein-carbohydrate interactions, as well as multi-subunit complexes. The universal library approach eliminates the need for separate prediction, cloning, and expression steps for each interaction pair, dramatically increasing throughput while maintaining operational simplicity.
3Productivity
If microfluidics-based platforms are used, then productivity is improved for identifying host proteins, but device complexity worsens due to complex microfluidic setup requirements
Solution Approach 1:
The patent extracts the core function of microfluidics-based platforms (high-throughput screening) and implements it using a simpler bead-based flow system. Instead of complex microfluidic channels and pumps, the invention uses magnetic beads displaying host proteins that can be manipulated with simple magnetic fields and flow systems. This extraction of the essential screening function eliminates complex microfluidic requirements while maintaining high throughput capability.
4Ease of manufacture
If conventional interaction screening methods are used, then ease of manufacture is improved, but loss of information worsens by missing secreted proteins involved in neutralization or exclusion from mucus layer
Solution Approach 1:
The patent changes the physical state and localization parameters of host secreted proteins by displaying them on the surface of phage particles. Secreted proteins that would normally be soluble and difficult to screen are instead presented in a stable, surface-displayed format on phage particles. This parameter change allows conventional screening methods to detect secreted proteins involved in neutralization or mucus exclusion while maintaining ease of manufacture through standard phage display protocols.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the high-throughput identification of host proteins interacting with microbes, facilitating the discovery of novel therapeutic targets and providing insights into microbial ecology and pathogenesis.
Implementation Method 1
A method using a display library of yeast cells expressing various proteins or peptides, such as nanobodies, to interact with microbial species
Implementation Method 2
contacting a first population of cells comprising one or more display molecules with a second population of cells comprising cells from at least one microbial species
Data Source
AI summary
The invention provides a BASEHIT screening method for identifying proteins that are involved in host-microbe interactions which may function as therapeutic targets.