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Quantitative ELISA for SERPINA4/kallistatin.
Biotechniques ( IF 2.2 ) Pub Date : 2021-09-16 , DOI: 10.2144/btn-2018-0194
Kurpad Nagaraj Shashidhar 1 , Venkata Lakshmaiah 2 , Chandrappa Muninarayana 3 , Krishna Sumanth Nallagangula 4
Affiliation  

Background: SERPINA4/kallistatin is a multifunctional protein expressed from the liver; its concentration in blood circulation reflects the degree of liver dysfunction and may serve as a diagnostic/prognostic biomarker for chronic liver diseases (CLD). Materials & methods: Antibodies specific for SERPINA4/kallistatin were used for the development of an enzyme-linked immunosorbent assay (ELISA). For correlative studies, blood samples from patients with cirrhotic liver and healthy patients were collected from R.L. Jalappa Hospital & Research Centre, Kolar. Results: Interference of other SERPINs was ruled out using western blot analysis. Quantitative ELISA was developed using monospecific antibodies as capture antibodies. Conclusion: The accuracy of the developed ELISA was determined by inter- and intra-assay precision. Linearity was defined using a spiked sample with serial dilutions. Reduced levels of SERPINA4/kallistatin were observed in patients with CLD compared with healthy controls.

中文翻译:

SERPINA4/kallistatin 的定量 ELISA。

背景: SERPINA4/kallistatin 是一种从肝脏表达的多功能蛋白。它在血液循环中的浓度反映了肝功能障碍的程度,可作为慢性肝病 (CLD) 的诊断/预后生物标志物。材料和方法:针对 SERPINA4/kallistatin 的特异性抗体用于开发酶联免疫吸附试验 (ELISA)。对于相关研究,来自 Kolar 的 RL Jalappa 医院和研究中心的肝硬化患者和健康患者的血液样本。结果:使用蛋白质印迹分析排除了其他 SERPIN 的干扰。使用单特异性抗体作为捕获抗体开发了定量 ELISA。结论:开发的 ELISA 的准确性由测定间和测定内的精度确定。使用连续稀释的加标样品定义线性。与健康对照组相比,在 CLD 患者中观察到 SERPINA4/kallistatin 水平降低。
更新日期:2021-09-16
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