Micro-Nanofluidic Chip for EV Cargo Loading
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Solution Overview
Problem
Existing methods for cargo loading of biological particles such as extracellular vesicles (EVs) face low efficiency and potential damage from electric fields, as well as issues like agglomeration and viability reduction, while micro- and nano-fluidic chips are lacking for this purpose.
Innovation Solution
A micro- and nano-fluidic chip design featuring alternately stacked nanochannel and microchannel array layers, allowing biological particles to be mechanically squeezed through nanochannels for cargo loading, eliminating the need for electric fields and reducing agglomeration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If electroporation method is used to load cargo into biological particles, then cargo-loading efficiency is improved, but external electric field causes excessive damage to biological particles and cargos
Solution Approach 1:
The patent replaces the electroporation method (which uses electrical fields) with a microfluidic squeezing method that uses controlled mechanical pressure. The microfluidic device applies pressure to biological particles to open pores in their membranes, allowing cargo loading without exposing particles to damaging external electric fields. This substitution of mechanical action for electrical action resolves the contradiction by maintaining high loading efficiency while eliminating electric field damage.
2Ease of operation
If incubation method is used to load cargo into biological particles, then operation simplicity is maintained, but cargo-loading efficiency is low
Solution Approach 1:
The patent introduces a microfluidic device as an intermediary tool between the simple incubation approach and the need for high loading efficiency. The device maintains operational simplicity by using a straightforward pressure-driven flow system, while simultaneously achieving high cargo-loading efficiency through controlled mechanical squeezing that effectively opens pores in biological particle membranes. This intermediary device bridges the gap between simplicity and effectiveness.
3Productivity
If electroporation method is used for cargo loading, then cargo-loading efficiency is improved, but agglomeration and viability reduction occur
Solution Approach 1:
The patent changes the fundamental parameter from electrical field strength to mechanical pressure magnitude and duration. By controlling the pressure parameters applied through the microfluidic device, the method achieves sufficient mechanical force to open pores for cargo loading while staying below the threshold that causes particle agglomeration or viability loss. This parameter optimization resolves the contradiction between loading efficiency and particle viability.
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 chip achieves high throughput and efficient cargo loading of biological particles with various cargos, avoiding damage and cost-effectively increasing efficiency compared to electroporation methods.
Implementation Method 1
the depth, width or diameter of the at least one nanochannel is less than or equal to the diameter of the biological particles, the membrane of the biological particles is mechanically squeezed in the at least one nanochannel so as to introduce pores
Data Source
Figure 1a~2c
Figure 3a~4c
Figure 5a~6c
AI summary
Provided is a micro- and nano-fluidic chip, including at least one nanochannel array layer and at least one microchannel array layer that are alternately stacked. The at least one nanochannel array layer includes nanochannels, the at least one microchannel array layer includes input units and/or output units. The input unit includes inlet microchannel arrays and inlets, and the output unit includes outlet microchannel arrays and outlets. The inlet microchannel array includes inlet microchannels, the outlet microchannel array includes outlet microchannels, and the inlet microchannels and the outlet microchannels are connected through the nanochannel. In this micro- and nano-fluidic chip, the nanochannel can effectively mechanically squeeze biological particles having a membrane structure such as extracellular vesicles (EVs) so as to introduce pores in the membrane of the biological particles such as EVs thereby realizing cargo-loading function, and can also effectively improve the loading efficiency of exogenous cargos, retain the biological viability of the biological particles, and increase the throughput of the cargo-loading process. In addition, a method for fabricating the above-mentioned micro- and nano-fluidic chip and a method for preparing cargo-loading biological particles are also provided.