This video demonstrates Microneutralization assays to measure neutralizing antibodies using Madin-Darby Canine Kidney cells overexpressing α2,6-linked sialic acid in human sera. This assay is often used for influenza human serology
Protocol
All influenza viruses should be handled according to appropriate biosafety level requirements (BSL-2 or higher) as defined in the Biosafety on Microbiological and Biomedical Laboratories (BMBL). 1. Preparation of Reagents and Starting Material Prepare Madine Darby Canine Kidney-α2,6-sial transferase (MDCK-SIAT1) cells and sterile cell culture medium Prepare the MDCK-SIAT1 cell culture medium using 500 mL of Dulb…
Representative Results
Figure 1. MDCK-SIAT1 cell culture. MDCK-SIAT1 cells. (A) MDCK-SIAT1 confluent monolayer (>100% confluency) for virus inoculation; (B) MDCK-SIAT1 cells at log phase with 75 – 95% confluency for TCID50 and MN assays. <img alt="Figure 2" src="/files/ftp_upload/21908/21908_Figure2.j…
Declarações
The authors have nothing to disclose.
Materials
Dulbecco's Modified Eagle Medium (DMEM) with high Glucose
Life Science
11965
A critical component of Sterile Cell Culture, Virus Propagation and Virus Diluent Media
Fetal bovine serum (FBS)
Hyclone
SH30070.03
Bovine Serum Albumin (BSA) Fraction V, Protase Free