Paper based immunosensing of ovarian cancer tumor protein CA 125 using novel nano-ink: A new platform for efficient diagnosis of cancer and biomedical analysis using microfluidic paper-based analytical devices (μPAD).

Ovarian cancer is the first and most important cause of malignancy death in women. Mucin 16 or MUC16 protein also known as carcinoma antigen 125 (CA 125) is the most commonly used glycoprotein for early stage diagnosis of ovarian cancer. In this work, a novel paper-based bio-device through hand writing of Ag/RGO (silver nanoparticles/reduced graphene oxide) nano-ink on the flexible paper substrate using pen-on-paper technology was developed. The prepared interface was used to the recognition of CA 125 protein in human biofluid. For this purpose, Ag/rGO nano-ink was synthesized by deposition of Ag nanoparticles onto graphene oxide sheets and the reduction of graphene oxide to rGO simultaneously. Conductivity and resistance of conductive lines were studied after drawing on photographic paper. Subsequently, to prepare a new and unique immuno-device, paper electrode modified by cysteamine caped gold nanoparticles (CysA/Au NPs) using electrochemical techniques. CysA is bonded by sulfur atoms with Au (CysA/Au NPs), and from the amine group with hydroxyl and carboxyl groups of Ag/RGO nano-ink deposited on the surface of paper-based electrodes (CysA/Au NPs/Ag-rGO). Then, anti-CA 125 antibody was immobilized on the electrode surface through Au NPs and CA 125 positively charged amine groups interaction. Atomic force microscopy, Transmission electron microscopy, Field emission scanning electron microscopy, and dynamic light scattering, were performed to identify the engineered immunosensor. Using chronoamperometry technique and under the optimized conditions, the low limit of quantitation (LLOQ) for the proposed immunoassay was recorded as 0.78 U/ml, which this evaluation was performed at highly linear range of 0.78-400 U/ml. The high sensitivity of the electrochemical immunosensor device is indicative of the ability of this immuno-device to detect early stages ovarian cancer.