Salehi Mohammad, Schneider Lilli, Ströbel Philipp, Marx Alexander, Packeisen Jens, Schlücker Sebastian
Department of Physics, University of Osnabrück, Barbarastr. 7, 49069 Osnabrück, Germany.
Nanoscale. 2014 Feb 21;6(4):2361-7. doi: 10.1039/c3nr05890e. Epub 2014 Jan 16.
SERS microscopy is a novel staining technique in immunohistochemistry, which is based on antibodies labeled with functionalized noble metal colloids called SERS labels or nanotags for optical detection. Conventional covalent bioconjugation of these SERS labels cannot prevent blocking of the antigen recognition sites of the antibody. We present a rational chemical design for SERS label-antibody conjugates which addresses this issue. Highly sensitive, silica-coated gold nanoparticle clusters as SERS labels are non-covalently conjugated to primary antibodies by using the chimeric protein A/G, which selectively recognizes the Fc part of antibodies and therefore prevents blocking of the antigen recognition sites. In proof-of-concept two-color imaging experiments for the co-localization of p63 and PSA on non-neoplastic prostate tissue FFPE specimens, we demonstrate the specificity and signal brightness of these rationally designed primary antibody-protein A/G-gold nanocluster conjugates.
表面增强拉曼散射显微镜术是免疫组织化学中的一种新型染色技术，它基于用称为表面增强拉曼散射标记物或纳米标签的功能化贵金属胶体标记的抗体进行光学检测。这些表面增强拉曼散射标记物的传统共价生物共轭不能防止抗体抗原识别位点的封闭。我们提出了一种针对表面增强拉曼散射标记物 - 抗体共轭物的合理化学设计，以解决这个问题。作为表面增强拉曼散射标记物的高灵敏度、二氧化硅包覆的金纳米颗粒簇通过使用嵌合蛋白A/G与一抗非共价共轭，该嵌合蛋白A/G选择性识别抗体的Fc部分，从而防止抗原识别位点的封闭。在用于p63和前列腺特异性抗原（PSA）在非肿瘤性前列腺组织福尔马林固定石蜡包埋（FFPE）标本中共定位的概念验证双色成像实验中，我们证明了这些合理设计的一抗 - 蛋白A/G - 金纳米簇共轭物的特异性和信号亮度。
