Biodistribution of Graphene Oxide NanosheetsUsing Post-Administration Labeling withOligonucleotide-Conjugated Gold Nanoparticles

• Main Authors: Lin, Jou-Yu, 林柔瑜
• Other Authors: Su, Cheng-Kuan
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/2j5ray

碩士 === 國立臺灣海洋大學 === 生命科學暨生物科技學系 === 107 === Graphene oxide (GO) has been considered as potential candidates for many biomedical applications due to their excellent aqueous processability, amphiphilicity, and surface functionalizability. However, to study the biodistribution of carbon-based nanomaterials (NMs) remains a technical barrier since those conventional methods relying on conjugating radiotracers or fluorescent probes onto these carbon-based NMs would change their surface properties and suffer the risks of tag detachment following circulation in bodies. To overcome these limitations, we employed the oligonucleotide-conjugated gold nanoparticles (AuNPs) to label these administrated GO in dissolved organs/tissues to allow quantification of GO through measuring gold signals by ICP-MS. To optimize the labeling of GO with oligonucleotide-conjugation AuNPs, the effects of the oligonucleotides (A20R20, A20R40, and A20R60, R: random sequence including A, T, C, G) as well as the incubation pH and time were evaluated. Also, an automatic sample pretreatment scheme utilizing a homemade minicolumn packed with C18 resins was optimized to efficiently separate these A20RX-AuNP-labeled GO from these residual A20RX-AuNPs. After this method’s optimization, it was found that the A20R20-AuNPs have the strongest binding affinity to label these administrated GO in biological samples with an equilibrium dissociation constant (Kd) of 36.0 fM, and the method’s detection limit reached 9.3 ag L-1. Moreover, our biodistribution results revealed that these intravenously administrated GO accumulated majorly in liver and spleen at 1 and 12 h post-administration, with apparent discrepancies in the GO concentraions acquired between the pre- and post-administration labeling strategies.