Formulation optimization of lyophilized aptamer-gold nanoparticles: Maintained colloidal stability and cellular uptake
Dalya Saidi,
Marya Obeidat,
Shrouq Alsotari,
Abed-Alqader Ibrahim,
Rula Al-Buqain,
Suha Wehaibi,
Dana A. Alqudah,
Hamdi Nsairat,
Walhan Alshaer,
Alaaldin M. Alkilany
Affiliations
Dalya Saidi
Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, Jordan University of Science and Technology, Irbid, 22110, Jordan
Marya Obeidat
Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, Jordan University of Science and Technology, Irbid, 22110, Jordan; Corresponding author. Department of Medical Laboratory Sciences, Faculty of Applied Medical Sciences, Jordan University of Science and Technology, Irbid, 22110, Jordan.
Shrouq Alsotari
Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan
Abed-Alqader Ibrahim
Department of Nanoscience, Joint School of Nanoscience and Nanoengineering, University of North Carolina at Greensboro, 2907 E. Gate City Blvd, Greensboro, NC, 27401, USA
Rula Al-Buqain
Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan
Suha Wehaibi
Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan
Dana A. Alqudah
Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan
Hamdi Nsairat
Pharmacological and Diagnostic Research Center, Faculty of Pharmacy, Al-Ahliyya Amman University, Amman, 19328, Jordan
Walhan Alshaer
Cell Therapy Center, The University of Jordan, Amman, 11942, Jordan
Alaaldin M. Alkilany
College of Pharmacy, QU Health, Qatar University, Doha, 2713, Qatar; Corresponding author. College of Pharmacy, Qatar University, Doha, 2713, Qatar.
Anti-nucleolin (NCL) aptamer AS1411 is the first anticancer aptamer tested in clinical trials. Gold nanoparticles (AuNP) have been widely exploited for various biomedical applications due to their unique functional properties. In this study, we evaluated the colloidal stability and targeting capacity of AS1411-funtionalized AuNP (AuNP/NCL-Apt) against MCF-7 breast cancer cell line before and after lyophilization. Trehalose, mannitol, and sucrose at various concentrations were evaluated to determine their cryoprotection effects. Our results indicate that sucrose at 10 % (w/v) exhibits the best cryoprotection effect and minimal AuNP/NCL-Apt aggregation as confirmed by UV–Vis spectroscopy and dynamic light scattering (DLS) measurements. Moreover, the lyophilized AuNP/NCL-Apt at optimized formulation maintained its targeting and cytotoxic functionality against MCF-7 cells as proven by the cellular uptake assays utilizing flow cytometry and confocal laser scanning microscopy (CLSM). Quantitative PCR (qPCR) analysis of nucleolin-target gene expression also confirmed the effectiveness of AuNP/NCL-Apt. This study highlights the importance of selecting the proper type and concentration of cryoprotectant in the typical nanoparticle lyophilization process and contributes to our understanding of the physical and biological properties of functionalized nanoparticles upon lyophilization.