Sørensen Morten Draeby, Gonzalez Dosal Regina, Jensen Kim Bak, Christensen Britta, Kølvraa Steen, Jensen Uffe Birk, Kristensen Peter
Department of Molecular Biology, University of Aarhus, Denmark.
J Biomed Biotechnol. 2009;2009:659219. doi: 10.1155/2009/659219. Epub 2009 Jul 14.
Invasive procedures for prenatal diagnosis are associated with increased risk of abortion; thus, development of noninvasive procedures would be beneficial. Based on the observation that embryonic nucleated red blood cell (NRBC) crosses the placenta and enters the circulation of pregnant women, the ability to identify such cell would allow development of such procedures. Identification of NRBCs in blood samples would be possible provided that specific antibodies are available. Here we have isolated recombinant antibodies using phage display. From the panel of antibody fragments specifically recognising epsilon-Hb, one was chosen for further characterization, DAb1. DAb1 binds to epsilon-Hb both in Western blots and immunocytochemistry. Several epsilon-Hb positive cells were detected in a blood sample taken as postchorionic villus sampling (CVS). To evaluate the sensitivity of the method, K562 cells (which express epsilon-Hb) were spiked in a blood sample followed by staining in solution and FACS analysis.
侵入性产前诊断程序与流产风险增加相关;因此,开发非侵入性程序将是有益的。基于胚胎有核红细胞(NRBC)穿过胎盘并进入孕妇循环的观察结果,识别此类细胞的能力将有助于开发此类程序。如果有特异性抗体,就有可能在血样中鉴定出NRBC。在这里,我们利用噬菌体展示技术分离出了重组抗体。从一组特异性识别ε-血红蛋白的抗体片段中,选择了一个进行进一步表征,即DAb1。DAb1在蛋白质印迹和免疫细胞化学中均能与ε-血红蛋白结合。在绒毛膜绒毛取样(CVS)后采集的一份血样中检测到了几个ε-血红蛋白阳性细胞。为了评估该方法的灵敏度,将K562细胞(表达ε-血红蛋白)加入血样中,然后进行溶液染色和荧光激活细胞分选分析。