Department of Pediatrics, Division of Infectious Diseases, University of California San Diego, La Jolla, CA 92093-0672, USA.
J Neurosci Methods. 2012 Jul 30;209(1):255-7. doi: 10.1016/j.jneumeth.2012.06.004. Epub 2012 Jun 21.
Mannose binding lectin (MBL) mediated complement pathway is an important constituent of innate immune response in several infections including neuroinflammatory and neurodegenerative diseases. Although there are Enzyme-Linked Immunosorbent Assays (ELISAs) for estimating MBL, MBL-associated serine protease-2 (MASP-2) and functional MBL-MASP-2 (fMBL) proteins for the plasma, serum and cell supernatants there are no established methods for their estimation in the cerebrospinal fluid (CSF). We developed sensitive ELISAs for the detection of MBL, fMBL and MASP-2 in the CSF. First, we adapted standard ELISAs for the detection of these proteins in the CSF. Second, we used a biotinyl-tyramide based horseradish peroxidase (HRP) signal amplification for the sensitive detection of these proteins in the CSF. In summary, using modified ELISA and biotinyl-tyramide based HRP signal amplification, we successfully detected MBL, fMBL and MASP-2 proteins in the CSF samples with high sensitivity and reproducibility.
甘露糖结合凝集素（MBL）介导的补体途径是包括神经炎症和神经退行性疾病在内的几种感染中先天免疫反应的重要组成部分。虽然有用于估计 MBL、MBL 相关丝氨酸蛋白酶-2（MASP-2）和功能性 MBL-MASP-2（fMBL）蛋白的酶联免疫吸附测定（ELISA），但尚无用于检测其在脑脊液（CSF）中的含量的既定方法。我们开发了用于检测 CSF 中 MBL、fMBL 和 MASP-2 的敏感 ELISA。首先，我们改编了标准 ELISA 以检测 CSF 中的这些蛋白质。其次，我们使用基于生物素化酪氨酸的辣根过氧化物酶（HRP）信号放大来灵敏地检测 CSF 中的这些蛋白质。总之，我们使用改良的 ELISA 和基于生物素化酪氨酸的 HRP 信号放大，成功地以高灵敏度和可重复性检测了 CSF 样品中的 MBL、fMBL 和 MASP-2 蛋白。
