新型磁性纳米粒子在杀伤性T细胞分离中的应用

THE APPLICATION OF NEW MAGNETIC NANOPARTICLES IN THE SEPARATION OF KILLER T CELLS

王丹阳

11849114

硕士

生物工程领域工程

葛东亮

2020-06-02

2020-07-20

哈尔滨工业大学

深圳

磁性纳米技术的应用促进了各个领域的发展，尤其是生物医学研究和临床领域。免疫细胞治疗作为一种新兴的癌症治疗手段，因其良好的治疗效果，近些年受到了越来越多的关注。在免疫细胞治疗的研究中，免疫细胞的分选显得尤为重要，分选技术会影响所需免疫细胞的质量。磁性纳米粒子具有磁响应强，良好的生物相容性，比表面积大等许多优点，科学家研究发现磁性纳米粒子在细胞分离中有着重要的应用价值。为更好的探究可应用于肿瘤杀伤性T淋巴细胞分离中的磁性纳米粒子，本课题利用实验室的新型磁性纳米平台，选用两种不同的磁性纳米粒子：羧基磁珠和氨基磁珠，进行CD8特异性抗体偶联实验，结合BCA微量蛋白浓度测定法探究两种磁性纳米粒子对抗体的偶联效果，发现在抗体添加量相同的条件下，氨基磁珠的偶联效果优于羧基磁珠。进一步通过抗体用量的梯度实验对氨基磁珠表面抗体的偶联量进行研究，结果显示在抗体与磁珠的比例为50 μg：3 mg时，氨基磁珠具有96.65%的最大偶联率。同时，利用Nicomp380/ZLS纳米粒度分析仪对磁珠与抗体的偶联反应前后进行粒度分析，结果显示免疫磁珠的平均粒径和多分散系数都有不同程度的增长，表明抗体的偶联会对免疫磁珠的分散性以及粒径产生影响，且在相同条件下氨基免疫磁珠的分散性优于羧基免疫磁珠，进一步证明了氨基磁珠比羧基磁珠更适合用于本课题研究。该课题旨在探究可以用于分离杀伤性T细胞的磁性纳米粒子，从磁性纳米粒子的选择入手，进行了抗体的偶联量以及免疫磁珠性质的分析，得到了对后续磁性纳米粒子在CD8+T淋巴细胞分离应用中的探究有参考价值的结论，为进行磁性纳米粒子参与的细胞分离以及CD8+T淋巴细胞在免疫细胞治疗方面的研究提供了理论依据。

The application of magnetic nanotechnology has promoted the development of various fields, especially in biomedical research and clinical fields. As a new cancer treatment method, immune cell therapy has attracted more and more attention in recent years due to its good therapeutic effect. In the research of immune cell therapy, the separation of immune cells is particularly important, and the sorting technology can influence the quality of the required immune cells. Magnetic nanoparticles have many advantages such as strong magnetic response, good biocompatibility, and large specific surface area. Scientists have found that magnetic nanoparticles have important potential application value in cell separation.In order to better study the magnetic nanoparticles which can be used in the separation of tumor killing T-lymphocytes, this project uses the new magnetic nanoplatform built in the laboratory, and selects two kinds of magnetic nanoparticles, carboxy magnetic beads and amino magnetic beads, for CD8 specific antibody coupling experiment. The coupling effect of this two kinds of magnetic nanoparticles on antibody is obtained by BCA method. It is found that the coupling effect of amino magnetic beads is better than that of carboxy magnetic beads under the condition of the same antibody dosage. Further, through the gradient experiment of antibody dosage, the coupling amount of the antibody on the surface of the amino magnetic bead is studied, the result shows that when the ratio of antibody to the magnetic bead is 50 μg: 3 mg, the maximum coupling rate of the amino magnetic beads is 96.65%. The experiment also uses Nicomp380/ZLS to analyze the particle size before and after the coupling reaction between the magnetic beads and the antibody. The results show that the average particle size and polydispersity of the immune magnetic beads have increased to varying degrees, indicating that the coupling of antibody can influence the dispersion and average particle size of immune beads, and it is found that the dispersion of the amino immune beads is better than that of the carboxyl immune beads under the same conditions, which further proves that amino magnetic beads are more suitable for the research of this project than carboxy magnetic beads.The purpose of this project is to study the magnetic nanoparticles that can be used to separate killer T cells, Through the study of the selection of magnetic nanoparticles, the amount of antibody coupling and the properties of immune magnetic beads, we can draw the conclusion that the magnetic nanoparticles have reference value in the study of CD8+T lymphocytes separation, and provides a theoretical basis for the subsequent study of cell separation with magnetic nanoparticles and CD8+T lymphocyte immunotherapy.

磁性纳米粒子 CD8特异性抗体 偶联反应 粒度分析

magnetic nanoparticles CD8 specific antibody coupling reaction particle size analysis

中文

联合培养

南方科技大学

王丹阳. 新型磁性纳米粒子在杀伤性T细胞分离中的应用[D]. 深圳. 哈尔滨工业大学,2020.