Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
Lab Chip. 2013 Mar 7;13(5):866-76. doi: 10.1039/c2lc41099k. Epub 2013 Jan 9.
The last decade has seen appreciable advancements in efforts towards increased portability of lab-on-a-chip devices by substituting microfluidics with molecular motor-based transportation. As of now, first proof-of-principle devices have analyzed protein mixtures of low complexity, such as target protein molecules in buffer solutions optimized for molecular motor performance. However, in a diagnostic work-up, lab-on-a-chip devices need to be compatible with complex biological samples. While it has been shown that such samples do not interfere with crucial steps in molecular diagnostics (for example antibody-antigen recognition), their effect on molecular motors is unknown. This critical and long overlooked issue is addressed here. In particular, we studied the effects of blood, cell lysates and solutions containing genomic DNA extracts on actomyosin and kinesin-microtubule-based transport, the two biomolecular motor systems that are most promising for lab-on-a-chip applications. We found that motor function is well preserved at defined dilutions of most of the investigated biological samples and demonstrated a molecular motor-driven label-free blood type test. Our results support the feasibility of molecular-motor driven nanodevices for diagnostic point-of-care applications and also demonstrate important constraints imposed by sample composition and device design that apply both to kinesin-microtubule and actomyosin driven applications.
过去十年,通过以基于分子马达的输送替代微流控技术,使得芯片实验室设备的便携性得到了显著提高。到目前为止,首批原理验证设备已经分析了低复杂度的蛋白质混合物,例如缓冲溶液中的靶蛋白分子,这些缓冲溶液是针对分子马达性能进行了优化的。然而,在诊断工作中,芯片实验室设备需要与复杂的生物样本兼容。虽然已经证明,此类样本不会干扰分子诊断中的关键步骤(例如抗体-抗原识别),但它们对分子马达的影响尚不清楚。本研究解决了这一关键但长期被忽视的问题。具体而言,我们研究了血液、细胞裂解物和含有基因组 DNA 提取物的溶液对肌球蛋白和驱动蛋白-微管的输送的影响,这两种生物分子马达系统是最有前途的用于芯片实验室应用的系统。我们发现,在大多数研究的生物样本的特定稀释度下,马达功能得到了很好的保留,并展示了一种基于分子马达的无标记血型测试。我们的研究结果支持了基于分子马达的纳米器件用于即时诊断应用的可行性,同时也证明了样本组成和器件设计所施加的重要限制,这些限制既适用于驱动蛋白-微管系统,也适用于肌球蛋白系统。