Microfluidic T Cell Sorting via Critical Post Array

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Solution Overview

Problem

Current autologous T cell therapies and CAR-T therapies face challenges in selecting T cells ex vivo to achieve the highest tumor killing potential, lacking effective methods for enriching T cells with desirable phenotypes.

Innovation Solution

A microfluidic device with a flow path featuring a first array of posts, characterized by a critical size of 4 to 7 microns, is used to enrich activated T lymphocytes by flowing a fluid sample through the device, allowing activated T cells to be separated based on size and tilt angle, enabling the identification and isolation of T cells specific to an antigen of interest.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional T cell selection methods are used, then T cells can be obtained for therapy, but the tumor killing potential is insufficient due to lack of effective enrichment for desirable phenotypes

Engineering Contradiction:
Improvetumor killing potentialVSAvoidenrichment efficiency
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The invention segments T cells based on their phenotypic characteristics, specifically separating activated T cells from resting T cells using a microfluidic device with size-based filtration. The device divides the cell population into different streams, allowing enrichment of activated T cells which have higher tumor killing potential.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

The invention changes the physical parameters of the selection process by using a microfluidic device with posts of specific dimensions (4-7 microns critical size) and specific tilt angles (45° to 135° relative to flow direction). These parameter changes enable size-based separation that enriches for activated T cells with desirable phenotypes.

Inventive Principle:
Principle #35Parameter changes

2Manufacturing precision

If T cells are sorted using the microfluidic device with posts of critical size 4-7 microns and tilt angle 45°-135°, then activated T lymphocytes are enriched, but the device complexity increases

Engineering Contradiction:
Improvecell separation precisionVSAvoidmicrofluidic device structure
Core Design Contradiction:
Manufacturing precisionVSDevice complexity

Solution Approach 1:

The invention replaces complex biological or chemical selection methods with a simplified mechanical filtration system. The microfluidic device uses purely mechanical size-based filtration through an array of posts, eliminating the need for complex antibodies, magnetic beads, or other sophisticated selection mechanisms while achieving high separation precision.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The invention uses a porous-like structure in the form of an array of posts with specific spacing and dimensions. The posts create gaps that act as size-selective pores, allowing smaller resting T cells to pass through while directing larger activated T cells into collection streams, achieving precise separation through the porous-structure analogy.

Inventive Principle:
Principle #31Porous materials

3Productivity

If the post array is configured with critical size 4-7 microns and specific tilt angles, then activated T cells are effectively separated, but the manufacturing precision requirements increase

Engineering Contradiction:
Improveenrichment yieldVSAvoidpost array fabrication tolerance
Core Design Contradiction:
ProductivityVSManufacturing precision

Solution Approach 1:

The invention defines specific parameter ranges for the post array (critical size 4-7 microns, tilt angles 45° to 135°) that optimize enrichment yield. By specifying ranges rather than single values, the design accommodates reasonable manufacturing variations while maintaining effective separation performance.

Inventive Principle:
Principle #35Parameter changes

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

The microfluidic device effectively enriches activated T lymphocytes, enhancing their tumor killing potential by selectively sorting and isolating T cells with specific antigen recognition capabilities, suitable for use as therapeutic agents.

Implementation Method 1

The first array is characterized by a critical size (D c ) of 4 microns to 7 microns. The posts of the first array are arranged in rows and columns... Adjacent posts in each respective column in the first array define gaps through which fluid can flow

Methodology Applied
Scientific EffectSize-based separation: Filter (physical)

Data Source

PatentEP3487510B1Sorting of t lymphocytes in a microfluidic device
Publication Date: 2023.07.19 BERKELEY LIGHTS INC
  • EP3487510B1 patent drawingFigure 1
  • EP3487510B1 patent drawingFigure 2A~2B
  • EP3487510B1 patent drawingFigure 2C~2D

AI summary

Methods of sorting T lymphocytes in a microfluidic device are provided. The methods can include flowing a fluid sample comprising T lymphocytes through a region of a microfluidic device that contains an array of posts. The array of posts can be configured to have a critical size (Dc) that separates activated T lymphocytes from nave T lymphocytes. Also provided are microfluidic devices having an array of posts configured to separate activated T lymphocytes from nave T lymphocytes, compositions enriched for T lymphocytes, particularly activated T lymphocytes that are known to be reactive to an antigen of interest, and methods of treating subjects suffering from a pathogenic disorder or cancer by administering such compositions.