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).

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