Monroe M R, Reddington A P, Cretich M, Chiari M, Little F, Ünlü M S
Biomedical Engineering Department BostonUniversity, Boston, Ma 02446, USA.
Annu Int Conf IEEE Eng Med Biol Soc. 2011;2011:478-81. doi: 10.1109/IEMBS.2011.6090069.
Variation of probe immobilization on microarrays hinders the ability to make high quality, assertive and statistically relevant conclusions needed in the healthcare setting. To address this problem, we have developed a calibrated, compact, inexpensive, multiplexed, dual modality point-of-care detection platform that calibrates and correlates surface probe density measured label-free to captured labeled secondary antibody, is independent of chip-to-chip variability, and improves upon existing diagnostic technology. We have identified four major technological advantages of our proposed platform: the capability to perform single spot analysis based on the fluorophore used for detection, a 10-fold gain in fluorescence signal due to optimized substrate, a calibrated, quantitative method that uses the combined fluorescent and label-free modalities to accurately measure the density of probe and bound target for a variety of systems, and a compact measurement platform offering reliable and rapid results at the doctor's office. Already, we have formulated over a 90% linear correlation between the amount of probe bound to surface and the resulting fluorescence of captured target for IgG, β-lactoglobulin, Ara h 1 peanut allergen, and Phl 5a Timothy grass allergen.
微阵列上探针固定的变化阻碍了在医疗环境中得出高质量、有说服力且具有统计学相关性结论的能力。为了解决这个问题，我们开发了一种经过校准的、紧凑的、廉价的、多重的、双模态即时检测平台，该平台可校准并关联无标记测量的表面探针密度与捕获的标记二抗，不受芯片间差异的影响，并对现有诊断技术进行了改进。我们已经确定了我们提出的平台的四个主要技术优势：能够基于用于检测的荧光团进行单点分析，由于优化的底物使荧光信号增强10倍，一种经过校准的定量方法，该方法使用荧光和无标记相结合的方式准确测量各种系统中探针和结合靶标的密度，以及一个紧凑的测量平台，可在医生办公室提供可靠且快速的结果。我们已经在与表面结合的探针量与捕获的IgG、β-乳球蛋白、Ara h 1花生过敏原和Phl 5a梯牧草过敏原靶标的荧光之间建立了超过90%的线性相关性。
