Pan Liu, Shuping Liu, Vidhi Dalal, Jerome Lane, Paolo Cravedi, Kirk Campbell, Andrea Angeletti, Xinfang Xie, Elisa Gessaroli, Eleonora Forte, Lorenzo Gallon, Jing Jin
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引用次数: 0
Abstract
Introduction: Recent studies discovered the prominent presence of anti-nephrin autoantibodies in minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS). However, widely different, and often unconventional autoantibody detection methods were used in these studies, making it challenging to standardize anti-nephrin antibody detection and quantification across different studies.
Methods: Here, we compare methods of conventional ELISA, immunoprecipitation (IP)- based on-beads ELISA, immunoprecipitation-Western blotting (IP-WB), and cell- and tissue-based immunofluorescence staining with two cohorts totaling 169 patients and control individuals.
Results: Different assay methods and antigen preparations led to method-specific false-positive and false-negative results. In general, high-quality antigens produced in human cells, combined with IP-based assays, yielded the most robust and reliable results. Among 63 and 24 samples from patients with FSGS or MCD, respectively, two patients with FSGS showed strong antibody signals in both ELISA-based assays and IP-WB, while approximately half of patients with MCD had weak signals detectable only by IP-WB.
Conclusion: These findings highlight the importance of standardizing antibody detection methods.
最近的研究发现，抗肾素自身抗体在微小改变病（MCD）和局灶节段性肾小球硬化（FSGS）中显著存在。然而，在这些研究中使用的自身抗体检测方法差异很大，而且通常是非常规的，这使得在不同的研究中标准化抗肾素抗体的检测和定量具有挑战性。方法：在这里，我们比较了常规ELISA、基于免疫沉淀（IP）的珠粒ELISA、免疫沉淀- western blotting （IP- wb）和基于细胞和组织的免疫荧光染色两种方法，共169例患者和对照个体。结果：不同的检测方法和抗原制备导致了方法特异性的假阳性和假阴性结果。一般来说，在人类细胞中产生的高质量抗原与基于ip的测定相结合，产生了最稳健和可靠的结果。在来自FSGS或MCD患者的63份和24份样本中，两名FSGS患者在基于elisa的检测和IP-WB中均显示出强抗体信号，而大约一半的MCD患者仅在IP-WB中检测到弱信号。结论：这些发现突出了规范抗体检测方法的重要性。
期刊介绍：
Kidney International (KI), the official journal of the International Society of Nephrology, is led by Dr. Pierre Ronco (Paris, France) and stands as one of nephrology's most cited and esteemed publications worldwide.
KI provides exceptional benefits for both readers and authors, featuring highly cited original articles, focused reviews, cutting-edge imaging techniques, and lively discussions on controversial topics.
The journal is dedicated to kidney research, serving researchers, clinical investigators, and practicing nephrologists.
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