Abstract:
Gold nanoparticles (AuNPs) with an estimate diameter of 25 nm have been synthesized and characterized using ultraviolet-visible spectroscopy (UV-Vis), dynamic light scattering (DLS) and Zeta potential measurements. AuNPs were coated with D2B, a monoclonal antibody recognizing an extracellular epitope of the human prostate specific membrane antigen (hPSMA). Binding of D2B to PSMA induces endocytosis of the latter, thereby marking PSMA as a docking site for the delivery of therapeutic agents.
To attain a stabilized and covalent bond between D2B and AuNPs, the sulthydryl group (SH) in a cysteine of the D2B was utilized. Binding of D2B to AuNPs-citrate colloidal solution caused a red shift with a higher wavelength of about 15 nm in the UV-Vis spectra. Furthermore, as a confirmation of the successful conjugation, DLS revealed an increase in both the AuNPs size from 25 to 63 rim and Zeta potential measurements for AuNPs-citrate from -45 mV to -23 mV. The cytotoxicity of D2B-AuNPs was assessed using the WST-1 cell proliferation assay and the agarose gel DNA fragmentation method.
Finally, specific delivery and binding of D2B-AuNPs was tested using flow cytometry and western blot.
Our results pave the way for further research using coated NPs as vehicles for drug delivery in both invitro and in vivo models.
Description:
M.S. -- Faculty of Natural and Applied Sciences, Notre Dame University, Louaize, 2018; "A thesis submitted in partial fulfillment of the requirement for the degree of Master of Science in Biology"; Includes bibliographical references (leaves 52-60).