Detection of Anti-SARS-CoV-2 IgG, IgM and IgA Using Advanced Flow Cytometry

Advance your COVID-19 research by obtaining a comprehensive profile of antibodies against SARS-CoV-2 using a multiplex and high throughput serological assay.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to persist due to constant mutation and evasion. Because development of a humoral immune response against this virus is critical for protection, there is an ongoing need to accurately assess the quantity and duration of antibody responses against SARS-CoV-2, particularly antibodies targeting the SARS-CoV-2 Spike protein receptor binding domain (RBD). The SARS-CoV-2 Spike RBD mediates viral entry into human cells by binding to the angiotensin-converting enzyme 2 (ACE2) and is the primary target of many COVID-19 vaccines currently in use. Antibodies against the SARS-CoV-2 Spike RBD generally have potent viral neutralizing activity and correlate with protective immunity against the virus. Serological testing to quantitate the three major isotypes (IgG, IgM and IgA) of antibodies specific for SARS-CoV-2 Spike RBD can be used to facilitate characterization and monitor the duration of antibody responses in both COVID-19 patients and vaccinated individuals, detect asymptomatic infections and survey past infection prevalence in a population. 

There are several methods for detecting anti-SARS-CoV-2 Spike RBD IgG, IgM or IgA antibodies, including a traditional enzyme-linked immunosorbent assay (ELISA), lateral flow immunoassay and flow cytometry. However, these methods have limitations:

  • Provide only qualitative results
  • Low sensitivity and specificity
  • Can be labor-intensive and may not support high-throughput applications
  • Do not allow multiplexing analysis of IgG, IgM and IgA antibodies
  • Require large amounts of reagents

The iQue® SARS-CoV-2 (IgG, IgM and IgA) kit provides a rapid, robust and sensitive serological assay for the measurement of IgG, IgM and IgA anti-SARS-CoV-2 Spike RBD antibodies in a single well of a 96 or 384-well plate with antibody detection levels in the pg/mL range. Combined detection of all three antibody isotypes  saves time and precious patient samples and provides a dynamic, comprehensive profile of humoral immune responses over time following natural infection with SARS-CoV-2 or immunization with COVID-19 vaccines. Accelerate research with the iQue® SARS-CoV-2 (IgG, IgM and IgA) kit!


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Application

Assay Concept

The iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit is a bead-based immunoassay designed for simultaneous detection of human IgG, IgM and IgA antibodies specific for SARS-CoV-2 Spike protein RBD produced by the immune system in response to viral infection or vaccination.

The iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit has been optimized for robust, multiplexed analysis on the iQue® platform with a VBR configuration. Qualitative and quantitative analysis of anti-SARS-CoV-2 Spike RBD antibodies from human blood samples (sera and plasma) is accomplished with a streamlined workflow and pre-set templates for ease of use. 

Basic workflow:

  • Add pre-diluted patient samples to 96 or 384-well plate
  • Add SARS-CoV-2 Spike RBD-coupled Beads to capture anti-SARS-CoV-2 Spike RBD antibodies 
  • Add SARS-CoV-2 Antibody Detection Cocktail containing fluorescently labeled anti-human IgG, IgM and IgA antibodies 
  • Analyze the plate on iQue® platform (VBR configuration).


Key Advantages

  • Obtain accurate assessment of immune response - Sensitive and simultaneous measurement of anti-SARS-CoV-2 Spike RBD IgG, IgM and IgA antibodies induced in response to natural infection or vaccination.

  • Maximize your productivity - Multiplexed assay enables high-throughput analysis of the three major isotypes (IgG, IgM and IgA) of antibodies specific for SARS-CoV-2 Spike RBD in a single well for comprehensive analysis of the humoral immune response.

  • Save precious samples - Streamlined workflow conserves precious samples - requires only a single 10 µL sample diluted at least 100-fold.

Obtain accurate assessment of immune response

Sensitive and simultaneous measurement of anti-SARS-CoV-2 Spike RBD IgG, IgM and IgA antibodies induced in response to natural infection or vaccination.


Figure 1.  Analysis of anti-SARS-CoV-2 Spike RBD IgG, IgM, and IgA antibodies in (A) serum from recovered COVID-19 patients (P1-4). Serum collected from two individuals prior to the pandemic were included as negative controls (Neg. 1-2).  (B) Analysis of serum collected from two individuals (VA & VB) prior to immunization (VA-0, VB-0), and again after the first (VA-1, VB-1) and second  injection (VA-2, VB-2)  with the BNT162b2 (Pfizer-BioNTech) COVID-19 vaccine. Serum samples were diluted 1:100 in Assay Diluent and then serially diluted 1:5 in triplicate to generate dilution curves.  

Maximize your productivity

Multiplexed assay enables high throughput analysis of the three major isotypes (IgG, IgM and IgA) of antibodies specific for SARS-CoV-2 Spike RBD in a single well for comprehensive analysis of the humoral immune response with real-time data analysis and novel visualization tools.


Figure 2.  Analysis of anti-SARS-CoV-2 Spike RBD IgG, IgM, and IgA antibodies in serum from recovered COVID-19 patients or individuals vaccinated with the BNT162b2 (Pfizer-BioNTech) vaccine.  Serum samples were diluted 1:100 in Assay Diluent and then serially diluted 1:3 in triplicate. Serum collected from two COVID-19 negative individuals were included as negative controls (Neg. Ctl). Each color represents triplicate dilution curves which were repeated in different quadrants of plate. (A.) Plate layout (B) Heat Map of anti-SARS-CoV-2  Spike RBD IgG concentrations per well (ng/mL). 

Save precious samples 

Streamlined workflow conserves precious samples.


Figure 3: iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit workflow. Easy to follow steps and low sample volumes needed (requires only a single 10 µL of plasma / serum diluted at least 100-fold).


 

Ordering Information

Platform: Compatible with iQue® 3 / iQue® Screener Plus - VBR Configurations

Product Name

Format

Cat. No.

iQue® SARS-CoV-2 (IgG, IgM and IgA) Kit




1 x 96 well

BA-97125

5 x 96 wells

BA-97126

1 x 384 wells

BA-97127

5 x 384 wells

BA-97128


Resources

Product Guide

iQue® SARS-CoV-2 (IgG, IgM, and IgG) Kit Product Guide

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iQue®️ Reagent Kits brochure cover

Brochure

Complete product listing of iQue® Reagents

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iQue®️ Advanced Flow Cytometry Platform brochure cover

Brochure

Drive your research forward with advanced flow cytometry – miniaturize sampling, multiplex bead and cell based assays,

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