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Method for preparing composite nano microsphere for enriching lung cancer cells

A technology of lung cancer cells and complex proteins, which is applied in the fields of polymer chemistry and cell biomedicine, can solve the problems of high false negative rate of lung cancer exfoliated cells and complex cell components, and achieve increased detection sample size, reduced costs, improved reliability and The effect of accuracy

Active Publication Date: 2012-02-08
南通得一科技咨询有限公司
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AI Technical Summary

Problems solved by technology

[0005] Therefore, the technical problem to be solved in the present invention is aimed at the complex cell components in sputum and the high false negative rate of lung cancer exfoliated cells in the detection of sputum, and provides a composite Fe 3 o 4 Magnetic nanoparticles and preparation method thereof, and a method for detecting lung cancer cells in sputum

Method used

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  • Method for preparing composite nano microsphere for enriching lung cancer cells
  • Method for preparing composite nano microsphere for enriching lung cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Fe 3 o 4 Preparation of nanoparticles

[0035]1) form ferric nitrate and ferrous nitrate in ethanol with the molar mass ratio of 4:1 to form a uniformly dispersed ethanol solution, wherein the concentration of iron ions is 0.5 to 1.0 mol / L; Epichlorohydrin whose mass is 10 times that of iron ions forms a sol, and forms a gel after standing; the prepared gel is aged and washed with ethanol and dried; followed by annealing at 400°C to obtain Fe 3 o 4 .

[0036] 2) After the reaction finishes, the Fe 3 o 4 Wash with deionized water and ethanol alternately until the pH value is 7. Finally, dry it under vacuum to get Fe 3 o 4 magnetic nanoparticles.

[0037] 3) Take 5g of Fe prepared above 3 o 4 Magnetic nanoparticles, placed in 20ml of deionized water, ultrasonic (1000kHz) dispersed for 30min and then centrifuged (or magnetically) separated to obtain nano-Fe 3 o 4 Precipitation A.

[0038] 4) Reset 5g of Precipitate A in 10ml of PBS buffer solution, ...

Embodiment 2

[0042] Example 2 Preparation of Composite Nanomagnetic Particles

[0043] 1. Take PEI-modified Fe 3 o 4 Nanoparticles 100mg were suspended in 10ml 0.01M pH7.2TBS to obtain a nanoparticle solution, in which PEI-modified Fe 3 o 4 The concentration of nanoparticles is about 3 × 10 13 pieces / ml.

[0044] 2. Preparation of complex proteins: Dissolve protein A (SPA) and protein B (rabbit anti-human CD44 and CK7) with 3ml 0.01M pH7.2TBS (both purchased from Yusen Biotechnology Co., Ltd.), mix well, and react at 4°C For 30 minutes, the protein A concentration was 10 mg / ml, and the protein B concentration was 1 mg / ml.

[0045] 3. Take 0.5ml of the above-mentioned AB composite protein solution and mix evenly with 10ml of the above-mentioned nanoparticle solution, and leave it at room temperature for 60 minutes to obtain a composite nanomagnetic particle solution.

Embodiment 3

[0046] Pathological detection of lung cancer cells in the sputum of embodiment 3

[0047] Sputum samples from 850 patients with lung cancer were randomly divided into two groups. Patients need to leave morning sputum for 3 consecutive days and send it to the hospital within 2 hours, otherwise, the sputum will deteriorate and the cells will autolyze, making it impossible to diagnose. Collect 5ml of the patient's sputum into a sputum box, add 0.5ml of alkaline lysate (containing 1% proteinase K, 10% NaOH in water) and mix well, keep at room temperature for 10min to fully liquefy the sputum.

[0048] The experimental group was subjected to lung cancer cell enrichment and liquid-based cytopathological detection. Add 1 / 10 volume of the composite nano-magnetic particle solution prepared in Example 2 to the above liquefied sputum, mix well, and let stand for 15 minutes. Centrifuge at 500 rpm for 5 minutes, take the precipitate and suspend it in preservation solution (10% aqueous me...

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Abstract

The invention discloses a composite Fe3O4 magnetic nano particle for combining lung cancer cells and a preparation method thereof. Polyimide is modified on the surface of the Fe3O4 magnetic nano particle, and composite protein which has capacity of combining the lung cancer cells is marked on the Fe3O4 magnetic nano particle, wherein the composite protein which has capacity of combining the lung cancer cells consists of staphylococcus A (SPA) protein and one or two of anti-CD44 antibody and anti-CK7 antibody. The invention also discloses a method for enriching and detecting the lung cancer cells in sputum. The method comprises the following steps of: a) collecting sputum, and fully liquefying the sputum; b) adding a composite nano magnetic particle solution and reacting; c) centrifuging reaction liquid to enrich cells in the reaction liquid; and suspending the obtained cells again, tabletting the suspension, dying, sealing and performing microscopic examination by using a thinprep cytology test (TCT) system. Compared with the conventional method, the method has the advantages that the positive rate of the lung cancer cells which is detected in the sputum is improved by more than 30 percent, the detection efficiency is improved obviously and detection cost is reduced to certain extent.

Description

technical field [0001] The invention belongs to the field of macromolecular chemistry and cell biomedicine, in particular to a compound Fe compound combined with lung cancer cells 3 o 4 Magnetic nanoparticles, a preparation method thereof, and a method for enriching and detecting lung cancer cells in sputum. Background technique [0002] Lung cancer ranks first among the top ten malignant tumors, and has the phenomenon of "three highs": high incidence, high mortality, and high rate of increase; it is a major disease affecting human health, and it has become one of the major medical problems in the 21st century alongside AIDS. With the current means of diagnosing lung cancer, once lung cancer is found and diagnosed, it is basically in the middle and late stages, and it is still difficult to obtain effective control. Therefore, realizing early diagnosis of lung cancer is an international medical problem that has not been solved for a long time to realize early treatment. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B82B1/00B82B3/00H01F1/36G01N1/34G01N33/50
Inventor 陈岗严彪李成魁李辉
Owner 南通得一科技咨询有限公司
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