Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Biochemistry and Molecular Engineering of Ministry of Education, College of Chemistry, Peking University, Beijing, China.
Talanta. 2011 Sep 15;85(3):1246-52. doi: 10.1016/j.talanta.2011.05.010. Epub 2011 Jun 12.
Orientedly bioconjugated core/shell Fe(3)O(4)@Au magnetic nanoparticles were synthesized for cell separation. The Fe(3)O(4)@Au magnetic nanoparticles were synthesized by reducing HAuCl(4) on the surfaces of Fe(3)O(4) nanoparticles, which were further characterized in detail by TEM, XRD and UV-vis spectra. Anti-CD3 monoclonal antibody was orientedly bioconjugated to the surface of Fe(3)O(4)@Au nanoparticles through affinity binding between the Fc portion of the antibody and protein A that covalently immobilized on the nanoparticles. The oriented immobilization method was performed to compare its efficiency for cell separation with the non-oriented one, in which the antibody was directly immobilized onto the carboxylated nanoparticle surface. Results showed that the orientedly bioconjugated Fe(3)O(4)@Au MNPs successfully pulled down CD3(+) T cells from the whole splenocytes with high efficiency of up to 98.4%, showing a more effective cell-capture nanostructure than that obtained by non-oriented strategy. This developed strategy for the synthesis and oriented bioconjugation of Fe(3)O(4)@Au MNPs provides an efficient tool for cell separation, and may be further applied to various fields of bioanalytical chemistry for diagnosis, affinity extraction and biosensor.
定向生物偶联的核/壳 Fe(3)O(4)@Au 磁性纳米粒子被合成用于细胞分离。通过在 Fe(3)O(4)纳米粒子表面还原 HAuCl(4)合成了 Fe(3)O(4)@Au 磁性纳米粒子，并用 TEM、XRD 和 UV-vis 光谱对其进行了详细的表征。抗 CD3 单克隆抗体通过抗体的 Fc 部分与共价固定在纳米粒子上的蛋白 A 之间的亲和力结合，定向生物偶联到 Fe(3)O(4)@Au 纳米粒子的表面。进行了定向固定化方法，以比较其与非定向方法的细胞分离效率，其中抗体直接固定在羧基化纳米粒子表面。结果表明，定向生物偶联的 Fe(3)O(4)@Au MNPs 能够从整个脾细胞中高效地拉下 CD3(+)T 细胞，效率高达 98.4%，显示出比非定向策略更有效的细胞捕获纳米结构。这种用于 Fe(3)O(4)@Au MNPs 的合成和定向生物偶联的开发策略为细胞分离提供了一种有效的工具，并可能进一步应用于生物分析化学的各个领域，用于诊断、亲和提取和生物传感器。
