Martínez-Moreno Julio Manuel, Llamas-Urbano Adrián, Barbarroja Nuria, Pérez-Sánchez Carlos
Cobiomic Bioscience SL. EBT UCO/IMIBIC, Cordoba, Spain.
Maimonides Institute for Research in Biomedicine of Cordoba (IMIBIC)/Reina Sofia University Hospital/University of Cordoba (UCO), Córdoba, Spain.
Methods Mol Biol. 2025;2929:129-142. doi: 10.1007/978-1-0716-4595-6_10.
The Proximity Extension Assay (PEA) is an innovative technology developed by Olink Proteomics that has revolutionized proteomics research. This method enables the simultaneous detection of hundreds of proteins using a minimal sample volume (1 microliter), combining the specificity of antibody-based immunoassays with the sensitivity of quantitative PCR (qPCR). The PEA process relies on the binding of two antibodies, each conjugated with unique DNA sequences, to a target protein. Upon proximity, these DNA sequences hybridize, forming a molecular barcode that is subsequently amplified by PCR, ensuring high specificity and minimizing cross-reactivity-an issue commonly encountered in multiplexed immunoassays. This chapter provides a detailed overview of the technical and methodological aspects of PEA, including the incubation, extension, amplification, and detection steps, as well as the quality control measures implemented in the Olink NPX Signature software for data analysis. This technology has been applied in more than 2000 peer-reviewed studies, demonstrating its potential for biomarker discovery in diagnostics, prognostics, and personalized medicine across various diseases. The integration of PEA into biomedical research continues to enhance the precision and reproducibility of proteomics studies.
邻近延伸分析（PEA）是由Olink蛋白质组学公司开发的一项创新技术，它彻底改变了蛋白质组学研究。该方法能够使用极少量样本（1微升）同时检测数百种蛋白质，将基于抗体的免疫分析的特异性与定量PCR（qPCR）的灵敏度相结合。PEA过程依赖于两种分别与独特DNA序列偶联的抗体与目标蛋白质的结合。当两种抗体靠近时，这些DNA序列杂交，形成一个分子条形码，随后通过PCR进行扩增，确保了高特异性并将多重免疫分析中常见的交叉反应降至最低。本章详细概述了PEA的技术和方法学方面，包括孵育、延伸、扩增和检测步骤，以及Olink NPX Signature软件中用于数据分析的质量控制措施。这项技术已应用于2000多项同行评审研究，证明了其在各种疾病的诊断、预后和个性化医疗中发现生物标志物的潜力。将PEA整合到生物医学研究中不断提高了蛋白质组学研究的准确性和可重复性。
