Kraft Fabio A, Baur Holger, Bommer Moritz, Latz Andreas, Fitschen-Oestern Stefanie, Fuchs Sabine, Gerken Martina
Integrated Systems and Photonics, Faculty of Engineering, Kiel University, Germany.
Kiel Nano, Surface and Interface Science KiNSIS, Kiel University, Germany.
Biomed Opt Express. 2023 Apr 26;14(5):2293-2310. doi: 10.1364/BOE.489138. eCollection 2023 May 1.
Direct detection of biomarkers from unpurified whole blood has been a challenge for label-free detection platforms, such as photonic crystal slabs (PCS). A wide range of measurement concepts for PCS exist, but exhibit technical limitations, which render them unsuitable for label-free biosensing with unfiltered whole blood. In this work, we single out the requirements for a label-free point-of-care setup based on PCS and present a wavelength selecting concept by angle tuning of an optical interference filter, which fulfills these requirements. We investigate the limit of detection (LOD) for bulk refractive index changes and obtain a value of 3.4 E-4 refractive index units (RIU). We demonstrate label-free multiplex detection for different types of immobilization entities, including aptamers, antigens, and simple proteins. For this multiplex setup we detect thrombin at a concentration of 6.3 µg/ml, antibodies of glutathione S-transferase (GST) diluted by a factor of 250, and streptavidin at a concentration of 33 µg/ml. In a first proof of principle experiment, we demonstrate the ability to detect immunoglobulins G (IgG) from unfiltered whole blood. These experiments are conducted directly in the hospital without temperature control of the photonic crystal transducer surface or the blood sample. We set the detected concentration levels into a medical frame of reference and point out possible applications.
对于诸如光子晶体平板(PCS)这样的无标记检测平台而言,直接从未经纯化的全血中检测生物标志物一直是一项挑战。存在多种用于PCS的测量概念,但它们都存在技术局限性,这使得它们不适用于对未过滤全血进行无标记生物传感。在这项工作中,我们明确了基于PCS的无标记即时检测装置的要求,并提出了一种通过光学干涉滤光片的角度调谐来选择波长的概念,该概念满足了这些要求。我们研究了体折射率变化的检测限(LOD),并获得了3.4×10⁻⁴折射率单位(RIU)的值。我们展示了针对不同类型固定化实体的无标记多重检测,包括适体、抗原和简单蛋白质。对于这种多重检测装置,我们检测到浓度为6.3μg/ml的凝血酶、稀释250倍的谷胱甘肽S-转移酶(GST)抗体以及浓度为33μg/ml的链霉亲和素。在第一个原理验证实验中,我们展示了从未经过滤的全血中检测免疫球蛋白G(IgG)的能力。这些实验是在医院直接进行的,没有对光子晶体换能器表面或血样进行温度控制。我们将检测到的浓度水平置于医学参考框架中,并指出了可能的应用。