Mudumba Sasi, de Alba Sophia, Romero Randy, Cherwien Carli, Wu Alice, Wang Jue, Gleeson Martin A, Iqbal Muzammil, Burlingame Rufus W
Genalyte, Inc., 10520 Wateridge Circle, San Diego, CA 92121, United States.
Genalyte, Inc., 10520 Wateridge Circle, San Diego, CA 92121, United States.
J Immunol Methods. 2017 Sep;448:34-43. doi: 10.1016/j.jim.2017.05.005. Epub 2017 May 17.
Photonic ring resonance is a property of light where in certain circumstances specific wavelengths are trapped in a ring resonator. Sensors based on silicon photonic ring resonators function by detecting the interaction between light circulating inside the sensor and matter deposited on the sensor surface. Binding of biological material results in a localized change in refractive index on the sensor surface, which affects the circulating optical field extending beyond the sensor boundary. That is, the resonant wavelength will change when the refractive index of the medium around the ring resonator changes. Ring resonators can be fabricated onto small silicon chips, allowing development of a miniature multiplex array of ring based biosensors. This paper describes the properties of such a system when responding to the refractive index changed in a simple and precise way by changing the ionic strength of the surrounding media, and in a more useful way by the binding of macromolecules to the surface above the resonators. Specifically, a capture immunoassay is described that measures the change of resonant wavelength as a patient serum sample with anti-SS-A autoantibodies is flowed over a chip spotted with SS-A antigen and amplified with anti-IgG. The technology has been miniaturized and etched into a 4×6mm silicon chip that can measure 32 different reactions in quadruplicate simultaneously. The variability between 128 rings on a chip as measured by 2M salt assays averaged 0.6% CV. The output of the assays is the average shift per cluster of 4 rings, and the assays averaged 0.5% CV between clusters. The variability between chips averaged 1.8%. Running the same array on multiple instruments showed that after some improvements to the wavelength referencing system, the upper boundary of variation was 3% between 13 different instruments. The immunoassay displayed about 2% higher variability than the salt assays. There are several outstanding features of this system. The amount of antigen used on the chip for each test is around 200 picograms, only a few microliters of sample is necessary, and the assays take <10min.
光子环共振是光的一种特性,在某些情况下,特定波长的光会被困在环形谐振器中。基于硅光子环谐振器的传感器通过检测传感器内部循环的光与沉积在传感器表面的物质之间的相互作用来发挥作用。生物材料的结合会导致传感器表面折射率的局部变化,这会影响延伸到传感器边界之外的循环光场。也就是说,当环形谐振器周围介质的折射率发生变化时,谐振波长也会改变。环形谐振器可以制造在小型硅芯片上,从而开发出基于环形的微型多重生物传感器阵列。本文描述了这样一个系统的特性,该系统通过改变周围介质的离子强度以简单而精确的方式响应折射率变化,并且通过大分子与谐振器上方表面的结合以更有用的方式响应折射率变化。具体而言,描述了一种捕获免疫测定法,当含有抗SS - A自身抗体的患者血清样本流过点有SS - A抗原并用抗IgG进行扩增的芯片时,该方法可测量谐振波长的变化。该技术已被小型化并蚀刻到一个4×6mm的硅芯片中,该芯片可以同时对32种不同反应进行四重测量。通过2M盐测定法测量,芯片上128个环之间的变异性平均为0.6%CV。测定的输出是每4个环的簇的平均偏移,并且簇之间的测定平均为0.5%CV。芯片之间的变异性平均为1.8%。在多台仪器上运行相同的阵列表明,对波长参考系统进行一些改进后,13台不同仪器之间的变化上限为3%。免疫测定法显示出比盐测定法高约2%的变异性。该系统有几个突出特点。每次测试在芯片上使用的抗原量约为200皮克,仅需要几微升的样品,并且测定时间<10分钟。