Dynamic Measurement of a Cancer Biomarker: Towards In Situ Application of a Fiber-Optic Ball Resonator Biosensor in CD44 Protein Detection

Abstract

The accuracy and efficacy of medical treatment would be greatly improved by the continuous and real-time monitoring of protein biomarkers. Identification of cancer biomarkers in patients with solid malignant tumors is receiving increasing attention. Existing techniques for detecting cancer proteins, such as the enzyme-linked immunosorbent assay, require a lot of work, are not multiplexed, and only allow for single-time point observations. In order to get one step closer to clinical usage, a dynamic platform for biosensing the cancer biomarker CD44 using a single-mode optical fiber-based ball resonator biosensor was designed, constructed and evaluated in this work. The main novelty of the work is an in-depth study of the capability of an in-house fabricated optical fiber biosensor for in situ detection of a cancer biomarker (CD44 protein) by conducting several types of experiments. The main results of the work are as follows: (1) Calibration of the fabricated fiber-optic ball resonator sensors in both static and dynamic conditions showed similar sensitivity to the refractive index change demonstrating its usefulness as a biosensing platform for dynamic measurements; (2) The fabricated sensors were shown to be insensitive to pressure changes further confirming their utility as an in situ sensor; (3) The sensor’s packaging and placement were optimized to create a better environment for the fabricated ball resonator’s performance in blood-mimicking environment; (4) Incubating increasing protein concentrations with antibody-functionalized sensor resulted in nearly instantaneous signal change indicating a femtomolar detection limit in a dynamic range from 7.1 aM to 16.7 nM; (5) The consistency of the obtained signal change was confirmed by repeatability studies; (6) Specificity experiments conducted under dynamic conditions demonstrated that the biosensors are highly selective to the targeted protein; (7) Surface morphology studies by AFM measurements further confirm the biosensor’s exceptional sensitivity by revealing a considerable shift in height but no change in surface roughness after detection. The biosensor’s ability to analyze clinically relevant proteins in real time with high sensitivity offers an advancement in the detection and monitoring of malignant tumors, hence improving patient diagnosis and health status surveillance.