Sensitive CSF ELISAs for the detection of MBL, MASP-2 and functional MBL/MASP-2.
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.
Department of Pediatrics, Division of Infectious Diseases, University of California San Diego, La Jolla, CA 92093-0672, USA.
Janet Y
JY
Kwok
Ryan M
RM
Augst
Denise Y
DY
Yu
Kumud K
KK
Singh
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D037601:Mannose-Binding Lectin / Q000134:cerebrospinal fluid*; D050606:Mannose-Binding Protein-Associated Serine Proteases / Q000134:cerebrospinal fluid*; D012680:Sensitivity and Specificity
D037601:Mannose-Binding Lectin; C105296:MASP2 protein, human; D050606:Mannose-Binding Protein-Associated Serine Proteases