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Advanced Western Blotting Solutions for SARS-CoV-2 Research, Serology Testing, and Vaccine Development

Bio-Techne is committed to supporting our customers including scientists who are working to combat the COVID-19 pandemic. Our next-generation western blotting solutions with Simple Western™ can help accelerate COVID-19 research and serology testing by enabling rapid, highly sensitive, and automated detection of proteins and serum antibodies involved in viral pathogenesis and host immune response.

The Simple Western Advantage in SARS-CoV-2 Research, Serology Testing, and Vaccine Development

Simple Western assays are automated, hands-free western blotting platforms that can be used for antigen-down serology assays for SARS-CoV-2. Simple Western systems like Jess™ and Abby™ can process 25 samples in 3 hours and offers the ability to use viral antigen ladders to measure serum antibodies reactive against multiple SARS-CoV-2 antigens simultaneously and provide information-rich views into differences in antibody binding patterns across patients. This approach also has promise as an orthogonal confirmatory test to validate results derived from lateral flow, ELISA or other immunoassay serology techniques.

Jess Instrument
Simple Western SARS CoV-2 Multi Antigen Serology Module WorkFlow

SARS-CoV-2 Serology Testing

How Simple Western Serology Assays Work

Serological assays on Simple Western load and separate a mixture of viral antigens according to their molecular weight. Simple Western is an open platform and users can utilize any antigens of relevance to their studies for serum antibody capture. Antigens can be recombinant viral proteins or viral lysates. After separation, viral antigens are immobilized using a proprietary immobilization chemistry. After immobilization, serum or plasma is introduced and IgG antibodies in the collected serum or plasma bind to the immobilized antigens. Anti-IgG, anti-IgM or other class- or isotype-specific secondary antibodies conjugated with HRP or a fluorescent tag can be used for detection.

Using a mixture of recombinant viral proteins or viral lysates in a serological assay may enable users to distinguish non-specific interactions and also provide rich insights into the relative binding capacity for several viral proteins in one run, providing increased confidence in results and a deeper understanding of the patient immune response.

 

The SARS-CoV-2 Multi-Antigen Serology Module

The SARS-CoV-2 Multi-Antigen Serology Module for Jess and Abby is an antigen down serology assay capable of detecting IgG serum antibodies reactive against 5 key viral antigens. Recombinant viral antigens are mixed and separated by size to create a ladder for capture of reactive serum antibodies. The module delivers the flexibility to utilize multiple viral antigens including the addition of proprietary vaccine antigens, providing more information about antibody binding profiles in each patient. Multiple patient samples can be processed in each 3-hour run, enabling scientists to characterize variability in the immune response to SARS-CoV-2.

SAR-CoV2 Multi Antigen Serology Module data set 3b

The Simple Western SARS-CoV-2 serology assay demonstrates differential patient reactivity to N, S1 RBD, S1 full length, S2 full length, and Spike (S1+S2) COVID-19 viral antigens using a Jess or Abby automated western blot system. A viral antigen ladder was used and tested with different PCR+ SARS-CoV-2 patient serum samples. SARS-CoV-2 positive serum samples were loaded onto a  Jess or Abby and human primary antibodies specific to each viral antigen were detected using an Anti-Human IgG HRP-conjugated secondary antibody (DM-005). The data is represented by quantitative analysis of relative peak areas.

The SARS-CoV-2 Multi-Antigen Serology Module provides all the antigens, controls, and diluents needed to develop your serological assay to detect serum antibodies reactive against recombinant Nucleocapsid protein (N), S1 Receptor Binding Domain protein (RBD), S1 subunit full length, S2 subunit full length, and Spike (S1+S2) viral antigens.

Learn how RAIN Incubator scientist, Shah A. K. Mohamed Bakhash, quickly compares human immune responses to five key viral antigens across a wide range of sample types, temporal serum draws, and dilutions.

SARS-CoV-2

COVID-19 Pathogenesis

Simple Western assays allow for highly sensitive and quantitative measurement of SARS-CoV-2 host proteins ACE2 and TMPRSS2 in human tissue and cell lysate samples. ACE2 binds the S protein of SARS-CoV-2 as an entry receptor. Cellular serine protease TMPRSS2 cleaves the Spike receptor at S1/S2 and S2’ sites to enable S2 mediated cell fusion and infection.

In this application note, Bio-Techne antibodies targeting ACE2 and TMPRSS2 were used in Simple Western assays that were highly sensitive over a large dynamic range, providing molecular weight information and even quantification of ACE2 and TMPRSS2 in human cells. These assays are valuable in the development of targeted therapies and to assess ACE2/TMPRSS2 density in various sample types as a measure of infection susceptibility.

a 3D COVID-19 spike protein for vaccine research
Nicolas Bazan thumbnail

From Your Peers

"It saves so much time! It takes away so much of the pipetting and manipulation error. You just load your samples and wait for results, and in the meantime, you can dedicate yourself to other activities."

- Nicolas G. Bazan, MD, PhD, Director of Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans

Dr. Bazan's laboratory used Simple Western's Jess to reveal a link between Elovanoids and ACE2 expression, presenting a new possible strategy to prevent and treat COVID-19. 

SARS-CoV-2 Vaccine Development

Simple Westerns are used routinely in vaccine development including in the following application areas:

  1. Serological assays to characterize immune response to a vaccine
  2. Characterizing vaccine proteins
  3. Monitoring vaccine component expression over time
  4. Quantitating vaccine manufacturing process intermediate contamination
Vaccine development process
Dr Ferrari Thumbnail

 

From Your Peers

Simple Western allowed me to screen quickly lots of samples with different antibodies while eliminating the manual washing and incubation steps.” 

- Dr. Matteo Ferrari, Postdoc, DIOSynVax, Department of Veterinary Medicine, Cambridge, United Kingdom

Quantikine ELISA Kits, most published

Learn how Simple Western was used to develop a COVID-19 vaccine which was published in Nature. Simple Western's ability to quantify large molecular weight proteins was harnessed to measure SARS-CoV-2 spike (S) protein expression in transduced cells.