PO.CH02.01 · 化学
解锁表面蛋白质组：利用Synlight-Rich与Synlight-Pure进行纳米尺度空间蛋白质组学以发现生物标志物和治疗靶点
Unlocking the surfaceome: Nanoscale spatial proteomics for biomarker and target discovery using Synlight-Rich and Synlight-Pure
该海报暂无可下载的资料
AACR 官方页面
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
细胞表面蛋白介导重要的信号传导、转运及细胞间相互作用，是关键的生物标志物和药物靶点类别。然而，由于现有蛋白质组学方法在空间精度和标记特异性方面存在局限，人类细胞表面蛋白质组的全面表征仍具挑战性。我们提出一套整合的工作流程，将Syncell的Microscoop® Mint平台与Synlight-Rich和Synlight-Pure试剂相结合，实现纳米尺度、无偏倚且具有极高特异性的表面蛋白质组发现。Synlight-Rich采用双光子光生物素化技术，在用户自定义的显微镜感兴趣区域（ROI）内以约350 nm的横向分辨率对蛋白进行共价标记，从而在高空间控制下对亚细胞区室进行深度蛋白质组学解析。被标记的蛋白通过Synpull试剂盒回收，并经LC-MS/MS分析。在此基础上，Synlight-Pure引入抗体介导的邻近标记，将生物素化限制在距靶结构约25–50 nm范围内。这种双重精度策略——图像引导与化学限定相结合——可实现对膜相关及相互作用邻近蛋白的选择性、高特异性富集。在HeLa细胞中，Microscoop结合Synlight-Rich鉴定出超过3,500种蛋白，其中包括超过1,000种已知的质膜蛋白，且超过50%的蛋白相较于未标记对照显示出≥1.5倍的富集（P < 0.05）。基因本体（Gene Ontology）分析显示，前200种富集蛋白中约55%定位于质膜区室。整合Synlight-Pure后，在同一富集组内膜特异性鉴定比例提升至约70%，凸显了标记特异性的显著改善，以及先前未表征的、参与受体和转运体介导信号传导的表面组分的发现。通过将空间蛋白质组学从微米尺度延伸至纳米尺度，Synlight-Rich与Synlight-Pure共同在单一工作流程中统一了无偏倚发现与靶向分子精度。该平台能够以无与伦比的特异性实现细胞表面蛋白质组的全面绘图，加速肿瘤学、神经退行性疾病及免疫学领域生物标志物和治疗靶点的鉴定。
查看英文原文 English abstract
Cell surface proteins mediate essential signaling, trafficking, and cell-cell interactions, representing key biomarker and drug target classes. Yet, comprehensive characterization of human cells surfaceome remains challenging due to the limited spatial precision and labeling specificity of existing proteomic methods. We present an integrated workflow using Syncell's Microscoop® Mint platform in combination with Synlight-Rich and Synlight-Pure reagents to achieve nanometer-scale, unbiased surfaceome discovery with exceptionally high specificity. Synlight-Rich employs two-photon photo-biotinylation to covalently tag proteins within user-defined microscopy regions of interest (ROI) at ~350 nm lateral resolution, enabling deep proteomic interrogation of subcellular domains with high spatial control. Labeled proteins are recovered via the Synpull kit and analyzed by LC-MS/MS. Building on this, Synlight-Pure introduces antibody-mediated proximity labeling, restricting biotinylation to within ~25-50 nm of the target structure. This dual-precision approach-image-guided and chemistry-confined-enables selective, high-specificity enrichment of membrane-associated and interaction-proximal proteins. In HeLa cells, Microscoop with Synlight-Rich identified >3,500 proteins, including >1,000 known plasma-membrane proteins, with >50% showing ≥1.5-fold enrichment (P < 0.05) over unlabeled controls. Gene Ontology analysis revealed that ~55% of the top 200 enriched proteins localized to plasma-membrane compartments. Integration of Synlight-Pure increased membrane-specific identifications to ~70% within the same enrichment group, highlighting substantial improvement in labeling specificity and the discovery of previously uncharacterized surface components involved in receptor- and transporter-mediated signaling. By extending spatial proteomics from the micrometer to nanometer scale, Synlight-Rich and Synlight-Pure together unify unbiased discovery and targeted molecular precision within a single workflow. This platform enables comprehensive cell surfaceome mapping with unparalleled specificity, accelerating biomarker and therapeutic target identification across oncology, neurodegeneration, and immunology.
利益披露 Disclosure
J. Liao,
Syncell Employment, Stock, Stock Option, Patent, Trademark, Copyright.
P. Chan,
Syncell Employment, Stock, Stock Option, Patent, Trademark, Copyright.
W. Chong,
Syncell Employment, Stock, Stock Option, Patent, Trademark, Copyright.
H. Chang,
Syncell Employment, Stock, Stock Option, Patent, Trademark, Copyright.
M. Oliveira,
Syncell Employment, Patent, Trademark, Copyright.
E. Huang,
Syncell Employment, Stock, Stock Option, Patent, Trademark, Copyright.
D. Dlugolenski,
Syncell Employment, Patent, Trademark, Copyright.