Fabrication method and application of an array sensor for transient recognition of drug-induced hk-2 cell damage
An array sensor and cell damage technology, applied in the field of biosensing, can solve the problems of cumbersome time-consuming, huge equipment, low sensitivity, etc., and achieve the effects of low cost, accurate typing and wide application range.
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[0036] like figure 1 Described; the preparation method of the array sensor for instantaneously identifying drug-induced HK-2 cell damage, the specific operation steps are as follows:
[0037] Step (1.1), preparation of polydopamine-polyethyleneimine (PDA-PEI) copolymer carrier: adding dopamine hydrochloride and polyethyleneimine to Tris-HCl buffer, stirring at room temperature 10-30°C in the dark After 2-8h, filter, place in a dialysis bag for dialysis for 24-36h; remove unreacted dopamine hydrochloride and polyethyleneimine, thereby obtaining a PDA-PEI copolymer carrier;
[0038] Specifically, 1. Dilute 100 μL Tris-HCl (1M, pH=7.4) buffer solution with ultrapure water to 10 mL for later use;
[0039] 2. Weigh 10 mg of dopamine hydrochloride (DA·HCl) and 10 mg of polyethyleneimine (PEI, M.W. 600Da), respectively, and add them to 10 mL of Tris-HCl buffer solution. Light stirring for 2-8h;
[0040] 3. Filter the above solution with a 0.22 μm cellulose ester membrane, then pla...
Embodiment 1
[0058] Construction of PDA-PEI / QDs sensor and verification of successful synthesis
[0059] 1. Preparation of PDA-PEI carrier
[0060] (1) Dilute 100 μL Tris-HCl (1M, pH=7.4) buffer solution with ultrapure water to 10 mL for later use;
[0061] (2), weigh 10mg of dopamine hydrochloride (DA·HCl) and 10mg of polyethyleneimine (PEI, M.W.600Da), respectively, add the two into 10mL Tris-HCl buffer solution, at room temperature 10-30 ℃ under a magnetic stirrer Stir in the dark for 2-8h;
[0062] (3), filter the above solution with 0.22 μm cellulose ester membrane, then place it in a dialysis bag (molecular weight cut-off 1000Da) for 24-36h dialysis to remove unreacted DA and PEI to obtain polydopamine-polyethyleneimine (PDA-PEI ) copolymer carrier;
[0063] During the preparation of PDA-PEI, the successful preparation of the PDA-PEI carrier was verified by the color change before and after the reaction, transmission electron microscopy, ultraviolet and infrared spectra.
[0064]...
Embodiment 2
[0078] Dosing (take isoniazid as an example), obtain IC by classical MTT method 50 value:
[0079] 1. The preparation process of the sensor is the same as that of Example 1;
[0080] 2. To study the cytotoxicity of isoniazid on HK-2 cells by MTT method. Include the following steps:
[0081] (1) Cell plate: HK-2 cells were plated at 4 × 10 per well 3 Density seeded into 96-well plates with 200 μL volume fraction of 12.5% FBS in NEAA-containing MEM basal medium per well (edge wells filled with sterile pH=7.4 PBS) at 37°C, 5% CO 2 Cultivated in incubator for 24h;
[0082] (2) Administration: after sucking off the culture medium, add 200 μL of isoniazid solution (with no isoniazid) of different concentration gradients (0, 0.1, 0.2, 0.6, 1.2, 2.4, 4.8, 9.6, 19.2, 38.4mM) respectively. FBS in MEM basal medium) was incubated at 37°C for 24h, and the cell morphology was observed under an inverted microscope;
[0083] (3) Give MTT: after the incubation is over, aspirate the m...
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