Emadi Elaheh, Bordbar Abdol-Khalegh, Nadri Hamid, Shams Ali, Taheri-Kafrani Asghar, Kalantar Seyed Mehdi
Department of Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services Yazd 8916978477 Iran
Department of Chemistry, University of Isfahan Isfahan 81746-73441 Iran.
RSC Adv. 2021 Sep 20;11(49):30990-31001. doi: 10.1039/d1ra05988b. eCollection 2021 Sep 14.
The development of an effective and noninvasive early method for obtaining fetal cells is crucial to prenatal screening. Despite proving the presence of fetal cells in the reproductive tract, their use is limited due to their inability to properly isolate them from maternal cells. Magnetic-activated cell sorting (MACS) is a simple technique to separate cells. The present study aimed to develop a MACS-based platform for the isolation of the HLA-G expressing trophoblast cells. For this purpose, first, the triazine functionalized MNPs were synthesized and characterized. Then, MNPs were directly and indirectly conjugated by the MEM-G/9 antibodies targeting HLA-G cells. The antibody amount on the surface of the nanoparticles was determined with the Bradford assay. The cell capture efficiency was also investigated. Various characterization methods confirmed the successful nanoparticle synthesis and antibody conjugation. The optimal initial antibody amount for the immobilization was about 20 μg and the optimal time was 3 h. The antibody-nanoparticles by the indirect method had better targeting and capture efficiency than the direct method. The MNPs indirectly conjugated with antibodies are an efficient tool for cell isolation and present considerable potential to be applied in biomedical fields.
开发一种有效且无创的获取胎儿细胞的早期方法对于产前筛查至关重要。尽管已证实在生殖道中存在胎儿细胞,但由于无法将其与母体细胞妥善分离,其应用受到限制。磁激活细胞分选(MACS)是一种简单的细胞分离技术。本研究旨在开发一个基于MACS的平台,用于分离表达HLA - G的滋养层细胞。为此,首先合成并表征了三嗪功能化的磁性纳米颗粒(MNPs)。然后,通过靶向HLA - G细胞的MEM - G/9抗体将MNPs直接和间接偶联。用Bradford测定法确定纳米颗粒表面的抗体量。还研究了细胞捕获效率。各种表征方法证实了纳米颗粒合成和抗体偶联的成功。固定的最佳初始抗体量约为20μg,最佳时间为3小时。间接法的抗体 - 纳米颗粒比直接法具有更好的靶向性和捕获效率。与抗体间接偶联的MNPs是细胞分离的有效工具,在生物医学领域具有相当大的应用潜力。