一种用于定量免疫调节蛋白的多重检测方法为免疫治疗临床试验中的相关性研究提供了支持。
A multiplexed assay for quantifying immunomodulatory proteins supports correlative studies in immunotherapy clinical trials.
作者信息
Whiteaker Jeffrey R, Zhao Lei, Schoenherr Regine M, Huang Dongqing, Lundeen Rachel A, Voytovich Ulianna, Kennedy Jacob J, Ivey Richard G, Lin Chenwei, Murillo Oscar D, Lorentzen Travis D, Colantonio Simona, Caceres Tessa W, Roberts Rhonda R, Knotts Joseph G, Reading Joshua J, Perry Candice D, Richardson Christopher W, Garcia-Buntley Sandra S, Bocik William, Hewitt Stephen M, Chowdhury Shrabanti, Vandermeer Jackie, Smith Stephen D, Gopal Ajay K, Ramchurren Nirasha, Fling Steven P, Wang Pei, Paulovich Amanda G
机构信息
Clinical Research Division, Fred Hutchinson Cancer Center, Seattle, WA, United States.
Cancer Research Technology Program, Antibody Characterization Lab, Frederick National Laboratory for Cancer Research, Frederick, MD, United States.
出版信息
Front Oncol. 2023 May 2;13:1168710. doi: 10.3389/fonc.2023.1168710. eCollection 2023.
INTRODUCTION
Immunotherapy is an effective treatment for a subset of cancer patients, and expanding the benefits of immunotherapy to all cancer patients will require predictive biomarkers of response and immune-related adverse events (irAEs). To support correlative studies in immunotherapy clinical trials, we are developing highly validated assays for quantifying immunomodulatory proteins in human biospecimens.
METHODS
Here, we developed a panel of novel monoclonal antibodies and incorporated them into a novel, multiplexed, immuno-multiple reaction monitoring mass spectrometry (MRM-MS)-based proteomic assay targeting 49 proteotypic peptides representing 43 immunomodulatory proteins.
RESULTS AND DISCUSSION
The multiplex assay was validated in human tissue and plasma matrices, where the linearity of quantification was >3 orders of magnitude with median interday CVs of 8.7% (tissue) and 10.1% (plasma). Proof-of-principle demonstration of the assay was conducted in plasma samples collected in clinical trials from lymphoma patients receiving an immune checkpoint inhibitor. We provide the assays and novel monoclonal antibodies as a publicly available resource for the biomedical community.
引言
免疫疗法是治疗部分癌症患者的有效方法，要将免疫疗法的益处扩展至所有癌症患者，需要有反应预测生物标志物和免疫相关不良事件（irAEs）。为支持免疫疗法临床试验中的相关研究，我们正在开发经过高度验证的检测方法，用于定量检测人体生物样本中的免疫调节蛋白。
方法
在此，我们开发了一组新型单克隆抗体，并将其纳入一种基于新型多重免疫多反应监测质谱（MRM-MS）的蛋白质组学检测方法中，该方法针对代表43种免疫调节蛋白的49种蛋白型肽段。
结果与讨论
该多重检测方法在人体组织和血浆基质中得到验证，定量线性范围>3个数量级，日间CV中位数在组织中为8.7%，在血浆中为10.1%。在接受免疫检查点抑制剂治疗的淋巴瘤患者临床试验收集的血浆样本中进行了该检测方法的原理验证。我们将这些检测方法和新型单克隆抗体作为生物医学界的公开可用资源提供。
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