Establishment of an immunoprecipitation-inductively coupled plasma mass spectrometry for detecting serum non-ceruloplasmin-bound copper and its clinical application
10.3760/cma.j.cn114452-20250625-00370
- VernacularTitle:免疫沉淀法-电感耦合等离子体质谱仪联用检测血清非铜蓝蛋白结合铜方法的建立及临床应用
- Author:
Junxia WU
1
;
Yiru XU
;
Qiang LU
;
Yaping FANG
;
Jianjian DONG
;
Chenchen XU
;
Zhichao HE
Author Information
1. 安徽中医药大学神经病学研究所，安徽中医药大学神经病学研究所附属医院检验科，合肥 230031
- Publication Type:Journal Article
- Keywords:
Immunoprecipitation;
Mass spectrometry;
Ceruloplasmin;
Copper;
Wilson′s disease;
Evaluation studies
- From:
Chinese Journal of Laboratory Medicine
2025;48(9):1215-1221
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.
