Sensitive CSF ELISAs for the detection of MBL, MASP-2 and functional MBL/MASP-2

被引:4
作者
Kwok, Janet Y. [1 ]
Augst, Ryan M. [1 ]
Yu, Denise Y. [1 ]
Singh, Kumud K. [1 ]
机构
[1] Univ Calif San Diego, Div Infect Dis, Dept Pediat, La Jolla, CA 92093 USA
关键词
Cerebrospinal fluid; MBL; Functional MBL; MASP-2; ELISA; MANNAN-BINDING LECTIN; CATALYZED REPORTER DEPOSITION; HUMAN SERUM; SIGNAL AMPLIFICATION; CEREBROSPINAL-FLUID; SERINE PROTEASE-2; BODY-FLUIDS; IMMUNOASSAYS; DEFICIENCY; ASSAY;
D O I
10.1016/j.jneumeth.2012.06.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. Published by Elsevier B.V.
引用
收藏
页码:255 / 257
页数:3
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