Center for Cancer Research, National Cancer Institute, National Institutes of Health, 10 Center Drive, Bethesda, Maryland 20892, USA.
Chembiochem. 2010 Feb 15;11(3):345-50. doi: 10.1002/cbic.200900532.
The human epidermal growth factor receptors, EGFR and HER2, are members of the EGFR family of cell-surface receptors/tyrosine kinases. EGFR- and HER2-positive cancers represent a more aggressive disease with greater likelihood of recurrence, poorer prognosis, and decreased survival rate, compared to EGFR- or HER2-negative cancers. The details of HER2 proto-oncogenic functions are not deeply understood, partially because of a restricted availability of tools for EGFR and HER2 detection (A. Sorkin and L. K. Goh, Exp. Cell Res. 2009, 315, 683-696). We have created photostable and relatively simple-to-produce imaging probes for in vitro staining of EGFR and HER2. These new reagents, called affiprobes, consist of a targeting moiety, a HER2- or EGFR-specific Affibody molecule, and a fluorescent moiety, mCherry (red) or EGFP (green). Our flow cytometry and confocal microscopy experiments demonstrated high specificity and signal/background ratio of affiprobes. Affiprobes are able to stain both live cells and frozen tumor xenograph sections. This type of optical probe can easily be extended for targeting other cell-surface antigens/ receptors.
人类表皮生长因子受体（EGFR）和人表皮生长因子受体 2（HER2）是细胞表面受体/酪氨酸激酶 EGFR 家族的成员。与 EGFR 或 HER2 阴性癌症相比，EGFR 和 HER2 阳性癌症具有更高的侵袭性、更大的复发可能性、更差的预后和更低的生存率。HER2 原癌基因功能的细节尚未被深入了解，部分原因是 EGFR 和 HER2 检测工具的可用性有限（A. Sorkin 和 L. K. Goh，Exp. Cell Res. 2009，315，683-696）。我们已经为 EGFR 和 HER2 的体外染色创建了光稳定且相对易于制备的成像探针。这些新的试剂称为亲和探针，由靶向部分、HER2 或 EGFR 特异性的 Affibody 分子和荧光部分（mCherry（红色）或 EGFP（绿色））组成。我们的流式细胞术和共聚焦显微镜实验证明了亲和探针具有高特异性和信号/背景比。亲和探针能够对活细胞和冷冻的肿瘤异种移植切片进行染色。这种类型的光学探针可以很容易地扩展到针对其他细胞表面抗原/受体。
