Integrated Display-Expression Vectors for High-Throughput Binder Screening

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Solution Overview

Problem

The conventional process of transferring inserts from display vectors to expression vectors for affinity binder identification and characterization is time-consuming, laborious, and has low throughput with unpredictable efficiency, typically ranging from 50% to 80%.

Innovation Solution

The design of vectors with judiciously engineered restriction enzyme sites and the use of combinations of restriction enzymes and DNA ligases facilitate high-throughput conversion from display to expression vectors, achieving conversion rates over 90% to 100%, thereby accelerating binder discovery and characterization processes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional subcloning process is used to transfer inserts from display vectors to expression vectors, then inserts can be expressed and characterized, but the process is time-consuming, laborious, and has low throughput

Engineering Contradiction:

Improvethroughput of binder characterizationVSAvoidtime required for subcloning process

Core Design Contradiction:

ProductivityVSLoss of time

Solution Approach 1:

The display vector and expression vector are merged into a single integrated vector design where the display function and expression function are combined in one molecule. This eliminates the need for separate subcloning steps and allows simultaneous display and expression in the same host system, dramatically increasing throughput and reducing time loss.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The vector is designed to perform multiple functions simultaneously: displaying the binder on the cell surface and expressing the binder for characterization. This multi-functional design consolidates what were previously separate processes into a single operation, resolving the contradiction between productivity and time loss.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Reliability

If conventional subcloning process is used, then inserts can be transferred to expression vectors, but the efficiency is low and unpredictable, varying from 50% to 80%

Engineering Contradiction:

Improveefficiency of subcloning processVSAvoidthroughput of binder characterization

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

By merging the display and expression functions into a single vector, the process eliminates the unreliable subcloning step entirely. The integrated vector ensures consistent and predictable transfer efficiency close to 100%, as the insert is already in the correct position and orientation for both display and expression without requiring additional cloning operations.

Inventive Principle:

Principle #5Merging (Combining)

3Ease of manufacture

If subcloning process is used to transfer inserts, then expression vectors can be generated, but the process is laborious and requires multiple steps

Engineering Contradiction:

Improveease of vector conversionVSAvoidcomplexity of conversion process

Core Design Contradiction:

Ease of manufactureVSDevice complexity

Solution Approach 1:

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The invention extracts and removes the complex subcloning steps from the workflow by designing a vector that inherently contains both display and expression capabilities. This simplification takes out the laborious intermediate steps and leaves only the essential function of generating the binder library, making the process much easier to manufacture and scale.

Inventive Principle:

Principle #2Taking out (Extraction)

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 significantly streamlines the process of affinity binder identification and characterization by achieving high conversion rates, reducing time and effort, and making it commercially desirable for affinity reagents discovery across various display platforms.

Implementation Method 1

The first and second restriction sites, when cleaved by corresponding restriction endonuclease thereto, produce compatible sticky ends

Methodology Applied

Scientific EffectRestriction enzyme cleavage: Enzyme

Implementation Method 2

employment of combinations of restriction enzymes and DNA ligases to facilitate the conversion

Methodology Applied

Scientific EffectDNA ligation: Chemical Bonding

Data Source

PatentUS20260071204A1Integrated system for library construction, affinity binder screening and expression thereof

Publication Date: 2026.03.12 ADAGENE INC

AI summary

A recombinant polynucleotide suitable for use in a display vector is provided. The recombinant polynucleotide includes from 5′ to 3′: a first nucleic acid sequence (or insert) encoding an amino acid sequence to be displayed on a surface; a first pre-selected restriction site; a second nucleic acid sequence encoding a surface peptide capable of being displayed on the surface; and a second pre-selected restriction site. Corresponding display vectors that can be converted into expression vectors in a high-throughput fashion, as well as methods of use thereof, are also provided.