Song Qing
Chemical and Biomolecular Engineering, New York University Polytechnic School of Engineering, 6 Metro Tech Center, Brooklyn, NY 11201, USA.
Sensors (Basel). 2015 Oct 16;15(10):26236-50. doi: 10.3390/s151026236.
Microengraving is a novel immunoassay for characterizing multiple protein secretions from single cells. During the immunoassay, characteristic diffusion and kinetic time scales and determine the time for molecular diffusion of proteins secreted from the activated single lymphocytes and subsequent binding onto the glass slide surface respectively. Our results demonstrate that molecular diffusion plays important roles in the early stage of protein adsorption dynamics which shifts to a kinetic controlled mechanism in the later stage. Similar dynamic pathways are observed for protein adsorption with significantly fast rates and rapid shifts in transport mechanisms when is increased a hundred times from 0.313 to 31.3. Theoretical adsorption isotherms follow the trend of experimentally obtained data. Adsorption isotherms indicate that amount of proteins secreted from individual cells and subsequently captured on a clean glass slide surface increases monotonically with time. Our study directly validates that protein secretion rates can be quantified by the microengraving immunoassay. This will enable us to apply microengraving immunoassays to quantify secretion rates from 10⁴-10⁵ single cells in parallel, screen antigen-specific cells with the highest secretion rate for clonal expansion and quantitatively reveal cellular heterogeneity within a small cell sample.
微刻蚀是一种用于表征单细胞多种蛋白质分泌的新型免疫测定法。在免疫测定过程中，特征扩散和动力学时间尺度分别决定了活化的单个淋巴细胞分泌的蛋白质分子扩散时间以及随后与载玻片表面结合的时间。我们的结果表明，分子扩散在蛋白质吸附动力学的早期阶段起着重要作用，而在后期则转变为动力学控制机制。当从0.313增加到31.3百倍时，观察到蛋白质吸附具有相似的动态途径，速率显著加快且运输机制迅速转变。理论吸附等温线遵循实验获得数据的趋势。吸附等温线表明，从单个细胞分泌并随后捕获在干净载玻片表面上的蛋白质数量随时间单调增加。我们的研究直接验证了蛋白质分泌速率可以通过微刻蚀免疫测定法进行量化。这将使我们能够应用微刻蚀免疫测定法并行量化10⁴ - 10⁵个单细胞的分泌速率，筛选出分泌速率最高的抗原特异性细胞进行克隆扩增，并定量揭示小细胞样本中的细胞异质性。
