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Detecting M-Protein via Mass Spectrometry and Affinity Beads: Enrichment With Mixed Kappa-Lambda Beads Enables Prompt Application in Clinical Laboratories.
Annals of Laboratory Medicine ( IF 4.0 ) Pub Date : 2024-08-20 , DOI: 10.3343/alm.2024.0039
Jikyo Lee 1, 2 , Jung Hoon Choi 3, 4 , Eun-Hee Kim 2 , Jihyun Im 2 , Heeyoun Hwang 3 , Seojin Yang 5 , Joon Hee Lee 1, 6 , Kyunghoon Lee 6 , Junghan Song 1, 6 , Seungman Park 7 , Sang Hoon Song 1, 2
Affiliation  

Background Detecting monoclonal protein (M-protein), a hallmark of plasma cell disorders, traditionally relies on methods such as protein electrophoresis, immune-electrophoresis, and immunofixation electrophoresis (IFE). Mass spectrometry (MS)-based methods, such as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray ionization-quadrupole time-of-flight (ESI-qTOF) MS, have emerged as sensitive methods. We explored the M-protein-detection efficacies of different MS techniques. Methods To isolate immunoglobulin and light chain proteins, six types of beads (IgG, IgA, IgM, kappa, lambda, and mixed kappa and lambda) were used to prepare samples along with CaptureSelect nanobody affinity beads (NBs). After purification, both MALDI-TOF MS and liquid chromatography coupled with Synapt G2 ESI-qTOF high-resolution MS analysis were performed. We purified 25 normal and 25 abnormal IFE samples using NBs and MALDI-TOF MS (NB-MALDI-TOF). Results Abnormal samples showed monoclonal peaks, whereas normal samples showed polyclonal peaks. The IgG and mixed kappa and lambda beads showed monoclonal peaks following the use of daratumumab (an IgG/kappa type of monoclonal antibody) with both MALDI-TOF and ESI-qTOF MS analysis. The limits of detection for MALDI-TOF MS and ESI-qTOF MS were established as 0.1 g/dL and 0.025 g/dL, respectively. NB-MALDI-TOF and IFE exhibited comparable sensitivity and specificity (92% and 92%, respectively). Conclusions NBs for M-protein detection, particularly with mixed kappa-lambda beads, identified monoclonal peaks with both MALDI-TOF and ESI-qTOF analyses. Qualitative analysis using MALDI-TOF yielded results comparable with that of IFE. NB-MALDI-TOF might be used as an alternative method to replace conventional tests (such as IFE) to detect M-protein with high sensitivity.

中文翻译:


通过质谱法和亲和珠检测 M 蛋白:使用混合 kappa-Lambda 珠进行富集可在临床实验室中迅速应用。



背景 检测单克隆蛋白 (M 蛋白) 是浆细胞疾病的标志,传统上依赖于蛋白质电泳、免疫电泳和免疫固定电泳 (IFE) 等方法。基于质谱 (MS) 的方法,例如基质辅助激光解吸/电离飞行时间 (MALDI-TOF) 和电喷雾电离四极杆飞行时间 (ESI-qTOF) MS,已成为敏感方法。我们探讨了不同 MS 技术的 M 蛋白检测功效。方法 为了分离免疫球蛋白和轻链蛋白,使用六种类型的微珠(IgG、IgA、IgM、kappa、lambda 以及混合 kappa 和 lambda)以及 CaptureSelect 纳米抗体亲和微珠 (NB) 制备样品。纯化后,进行 MALDI-TOF MS 和液相色谱联用 Synapt G2 ESI-qTOF 高分辨率 MS 分析。我们使用 NBs 和 MALDI-TOF MS (NB-MALDI-TOF) 纯化了 25 个正常和 25 个异常 IFE 样品。结果 异常样品显示单克隆峰,而正常样品显示多克隆峰。使用 daratumumab (一种 IgG/kappa 类型的单克隆抗体) 进行 MALDI-TOF 和 ESI-qTOF MS 分析后,IgG 以及 kappa 和 lambda 混合珠子显示单克隆峰。MALDI-TOF MS 和 ESI-qTOF MS 的检测限分别为 0.1 g/dL 和 0.025 g/dL。NB-MALDI-TOF 和 IFE 表现出相当的敏感性和特异性 (分别为 92% 和 92%)。结论 M 蛋白检测的 NBs,特别是使用混合 kappa-lambda 微珠,通过 MALDI-TOF 和 ESI-qTOF 分析鉴定了单克隆峰。使用 MALDI-TOF 的定性分析产生的结果与 IFE 的结果相当。 NB-MALDI-TOF 可用作替代常规测试(如 IFE)的替代方法,以高灵敏度检测 M 蛋白。
更新日期:2024-08-20
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