Using a Sandwich ELISA to Determine the Immunogenic Glycoprotein Content in a Vaccine
Published: April 30, 2024
Abstract
Source: Jallet, C. et al., In Vitro ELISA Test to Evaluate Rabies Vaccine Potency. J. Vis. Exp. (2020)
This video showcases an indirect enzyme-linked immunosorbent assay or ELISA sandwich immunocapture assay designed to assess immunogenic glycoprotein levels in vaccines. The assay plate, containing glycoprotein-specific antibodies, is incubated with serially diluted vaccine samples, including reference and test vaccines, followed by immunoassay to quantify glycoprotein content in the test vaccine.
Protocol
1. Security precautions NOTE: This method is applicable to both live rabies virus (RABV) and inactivated vaccine. Use good Laboratory Practice and Safety procedures. Wear adequate Personal Protection Equipment (PPE) including disposable coat, gloves, masks, glasses, etc. When the live virus is titrated, use a class II biological safety cabinet. Consider any material in contact with the samples (reagents, washing solu…
Divulgaciones
The authors have nothing to disclose.
Materials
| Class II Biological Safety Cabinet | ThermoFisher Scientific | 10445753 | If titrating live virus |
| Clear Flat-Bottom Immuno Nonsterile 96-Well Plates, 400 µL, MAXISORP | ThermoFisher Scientific | 439454 | Good for binding to the loaded antibody |
| Equip Labo Polypropylene Laboratory Fume Hood | ThermoFisher Scientific | 12576606 | For the preparation of sulfuric acid |
| Immunology Plate Strong Adsorption MAXISORP Flat Bottom Well F96 |
Dutscher | 55303 | Good for binding to the loaded antibody |
| Microplate Sealing Tape(100 sheets) | ThermoFisher Scientific | 15036 | |
| Microplate single mode reader Sunrise | TECAN | ||
| Microplate shaker-incubator | Dutscher | 441504 | |
| Microplate washer Wellwash | ThermoFisher Scientific | 5165000 | |
| Multichannel pipette (30-300 µL) 12 channels | ThermoFisher Scientific | 4661180N | |
| Single Channel pipettes (Kit 2 : Finnpipettes F2 0.2-2 μL micro, 2-20 μL, 20-200 μL & 100-1000 μL) | ThermoFisher Scientific | 4700880 |
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Using a Sandwich ELISA to Determine the Immunogenic Glycoprotein Content in a Vaccine. J. Vis. Exp. (Pending Publication), e22118, doi: (2024).