Cancer Research Center, Medical College, Xiamen University, Xiamen, China.
Department of Pediatrics, Xiang'an Hospital of Xiamen University, Xiamen, China.
J Clin Lab Anal. 2019 May;33(4):e22850. doi: 10.1002/jcla.22850. Epub 2019 Feb 13.
Neuropilins (Nrps) are a new type of broad-spectrum tumor marker. Currently, a method for accurate simultaneous quantification of Nrps is not available. We aimed to develop a bead-based and duplexed flow cytometric assay that could be used for accurate and simultaneous quantification of Nrp1 and Nrp2 for scientific research or clinical diagnosis.
We coupled anti-human Nrp1-11# mAb and anti-human Nrp2-C3 mAb to magnetic beads 18# and 25#, respectively. Capturing antibodies and detecting antibodies were then combined to detect Nrps by a bead-based Luminex assay, which was subsequently applied to quantify Nrps in clinical serum samples.
The results showed that the detection value of Nrps ranged from 10 to 100 000 pg/mL for Nrp1 and from 25 to 100 000 pg/mL for Nrp2. The detection sensitivity reached 10 pg/mL for Nrp1 and 24.8 pg/mL for Nrp2. Intra-assay variances ranged from 1.0% to 2.6% for Nrp1 and from 2.9% to 4.0% for Nrp2, and interassay variances ranged from 1.5% to 6.4% for Nrp1 and from 4.2% to 8.1% for Nrp2. The Nrp1 and Nrp2 recoveries were 96.6%-103.6% and 95.6%-102.3%, respectively. Irrelevant antigens had no interference in the paired-detection system, and the mean fluorescence intensity (MFI) values were stable for months.
A bead-based, duplexed flow cytometric assay (xMAP technology) was developed to detect Nrp1 and Nrp2. The assay provided rapid, high-throughput results and was much more sensitive, specific, reproducible, and stable than existing assays. In addition, this assay could be applied in early-stage cancer screening, tumor malignancy analysis, and prognosis assessment.
神经纤毛蛋白（Nrps）是一种新型广谱肿瘤标志物。目前，尚无准确同时定量检测 Nrps 的方法。我们旨在开发一种基于微球的双流式细胞术检测方法，用于准确、同时定量检测 Nrp1 和 Nrp2，以用于科研或临床诊断。
我们将抗人 Nrp1-11# mAb 和抗人 Nrp2-C3 mAb 分别偶联到磁珠 18#和 25#上。然后将捕获抗体和检测抗体组合在一起，通过基于微球的 Luminex 检测法检测 Nrps，随后将其应用于临床血清样本中 Nrps 的定量检测。
结果表明，Nrps 的检测值范围为 Nrp1 的 10 至 100000pg/mL，Nrp2 的 25 至 100000pg/mL。检测灵敏度达到 Nrp1 的 10pg/mL 和 Nrp2 的 24.8pg/mL。Nrp1 的批内变异系数范围为 1.0%至 2.6%，Nrp2 的批内变异系数范围为 2.9%至 4.0%；Nrp1 的批间变异系数范围为 1.5%至 6.4%，Nrp2 的批间变异系数范围为 4.2%至 8.1%。Nrp1 和 Nrp2 的回收率分别为 96.6%至 103.6%和 95.6%至 102.3%。无关抗原对配对检测系统无干扰，平均荧光强度（MFI）值稳定数月。
我们开发了一种基于微球的双流式细胞术检测方法（xMAP 技术）用于检测 Nrp1 和 Nrp2。该检测方法快速、高通量，与现有检测方法相比，灵敏度更高、特异性更强、重现性和稳定性更好。此外，该检测方法可用于早期癌症筛查、肿瘤恶性分析和预后评估。
