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Biological chip for delivering target molecules to cells and preparation method and application thereof

A biochip and molecular technology, applied in biochemical equipment and methods, bioreactor/fermenter for specific purposes, bioreactor/fermenter combination, etc. Bending and other problems, to achieve the effect of high cell safety, high delivery efficiency, and high throughput

Active Publication Date: 2019-11-15
北京载愈生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many technical problems in the 3D NEP silicon-based chip used in in vivo gene transfection experiments: (1) the silicon chip for constructing nanoscale channels cannot be bent, and is poorly compatible with the curvature of the skin; (2) it is fragile , the reuse efficiency is poor; (3) the gene solution cannot be packaged, and a large amount of biological reagents need to be consumed; (4) the cost of silicon nanochips prepared by projection lithography and DRIE technology is very high, and it is not easy to carry out large-scale in vivo applications

Method used

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  • Biological chip for delivering target molecules to cells and preparation method and application thereof
  • Biological chip for delivering target molecules to cells and preparation method and application thereof
  • Biological chip for delivering target molecules to cells and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Biochip

[0054] Such as figure 1 As shown, in this embodiment, the biochip comprises (1) a molecule delivery region; (2) an NFC module; and optionally (3) an impedance sensor. The molecular delivery area, NFC module and impedance sensor can be directly built on the flexible substrate, which can be bent and attached directly to the skin (after exfoliation and dead skin)( figure 2 ).

[0055] The material of the flexible substrate can be selected from flexible polymer materials such as polyimide (PI), polyethylene terephthalate (PET), polycarbonate (PC) or polydimethylsiloxane (PDMS) etc., where the polyimide can be thermoplastic polyimide or a material containing polyimide, for example: APICAL TM ; apoly-oxydiphenylene-pyromellitic imide, or KAPTON TM ; Biphenyltetracarboxylic dianhydride (BPDA) polymer, or VTECPI TM ;NORTONTH TM ;KAPTREX TM Wait.

[0056] (1) Molecular delivery area

[0057] The molecular delivery area includes microelectrode arra...

Embodiment 2

[0072] Example 2 Process scheme for preparing biochip

[0073] Such as Figure 6 As shown, in some embodiments of the present invention, the preparation process of the biochip includes the following steps:

[0074] (a) Coating on a substrate 1 (e.g. a silicon wafer) an adhesive layer 2 for bonding flexible substrates, such as colloidal polydimethylsiloxane (PDMS) comprising a polymeric organosilicon compound or equivalent ;

[0075] (b) A flexible substrate 3 such as a polyimide film is bonded to the adhesive layer 2, followed by heat treatment. For example, the bonding time is about 1 hour to 3 hours when the vacuum is lower than ~1kPa, the temperature is about 60°C to 80°C;

[0076] (c) Coating a photoresist 4 (for example, EPI 680TM (Everlight Chemical, Taiwan, China)) on the flexible substrate 3 by means of a homogenizer. The thickness of the photoresist is about 2 μm to 4 μm, and patterned to form microelectrodes, NFC circuits, Ultra-thin magnetic helical antenna, imp...

Embodiment 3

[0084] Example 3 Delivery of dacarbazine (dacarbazine) to cells using a biochip

[0085] In the dacarbazine delivery experiment, malignant melanoma cells (such as A375 cells) were cultured and placed on the nanoporous membrane of the chip. To prevent cell leakage, a cavity is constructed around the nanoporous membrane using PDMS, in which the cell fluid and cells rest. After overnight incubation, the cells adhered tightly to the nanopores.

[0086] During the electroporation experiment, the dacarbazine solution with a concentration of 5 μM was passed through the chip inlet to fill the entire microchannel array. Since this experiment is an isolated cell experiment, a platinum electrode was used as the bottom electrode in the cell solution, and the platinum electrode and the microelectrode array were respectively connected to a commercial electrical signal generator (for example, PowerStation of BioRad Gene Pulser, etc.). The square wave voltage signal used for electroporation...

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Abstract

The invention discloses a biologic chip for delivering target molecules to cells and a preparation method and application thereof. The biologic chip comprises a molecule delivery region and an NFC module; the molecule delivery region and the NFC module are arranged on a flexible substrate, the molecule delivery region comprises a micro-electrode array formed on the flexible substrate, a micro-channel array formed on the micro-electrode array and a nanometer porous film covering the micro-channel array, the NFC module comprises an NFC circuit and an antenna, the antenna is connected with an NFCcircuit, one end of the NFC circuit is connected with the micro-electrode array, and the other end of the NFC circuit is connected with a needle electrode on the flexible substrate.

Description

technical field [0001] The invention belongs to the technical field of biochips, in particular to a biomedical nanochip that can be directly applied to the skin and directly deliver target molecules into cells, as well as its preparation method and application. Background technique [0002] Gene therapy is currently one of the effective ways to treat skin cancer. This method directly introduces exogenous gene molecules into skin cells, changes the expression of intracellular target genes, and finally achieves the purpose of cancer cell death; at the same time, gene molecules do not affect normal skin cells. In the normal cellular environment, these exogenous molecules cannot directly cross the cell membrane, enter the cell and produce effects. Therefore, the core issue in the process of gene therapy is how to quickly, efficiently and safely introduce gene molecules directly into target skin cells in vivo. [0003] Common in vivo gene transfection techniques include viral v...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12M1/00A61M37/00C12N15/90
CPCA61M37/00A61M2037/0007C12N15/907C12Q1/6837
Inventor 常凌乾郝永存常洪龙
Owner 北京载愈生物科技有限公司
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