Craig Gary A, Allen Peter J, Mason Michael D
Department of Chemical and Biological Engineering, University of Maine, Orono, ME, USA.
Methods Mol Biol. 2010;624:177-93. doi: 10.1007/978-1-60761-609-2_12.
This chapter describes the methodology by which mAb-F19-conjugated gold nanoparticles were prepared and used to label human pancreatic adenocarcinoma. Specifically, gold nanoparticles were coated with dithiol bearing hetero-bifunctional PEG (polyethylene glycol), and cancer-specific mAb F19 was attached by means of NHS-EDC coupling chemistry taking advantage of a carboxylic acid group on the heterobifunctional PEG. These conjugates were completely stable and were characterized by a variety of methods, including UV-Vis absorbance spectrometry, darkfield microscopy, DLS (dynamic light scattering), TEM (transmission electron microscopy), SEC (size-exclusion chromatography), and confocal microscopy. Nanoparticle bioconjugates were used to label sections of healthy and cancerous human pancreatic tissue. Labeled tissue sections were examined by darkfield microscopy and indicate that these nanoparticle bioconjugates may selectively bind to cancerous tissue and provide a means of optical contrast.
本章描述了制备单克隆抗体F19偶联金纳米颗粒并将其用于标记人胰腺腺癌的方法。具体而言，用带有杂双功能聚乙二醇（PEG）的二硫醇包覆金纳米颗粒，并利用杂双功能PEG上的羧酸基团通过NHS-EDC偶联化学连接癌症特异性单克隆抗体F19。这些偶联物完全稳定，并通过多种方法进行了表征，包括紫外可见吸收光谱法、暗场显微镜、动态光散射（DLS）、透射电子显微镜（TEM）、尺寸排阻色谱（SEC）和共聚焦显微镜。纳米颗粒生物偶联物用于标记健康和癌性人胰腺组织切片。通过暗场显微镜检查标记的组织切片，结果表明这些纳米颗粒生物偶联物可能选择性地结合癌组织，并提供光学对比手段。
