Rapid and accurate agglutination-based testing for SARS-CoV-2 antibodies

Esmail, Sally; Knauer, Michael J.; Abdoh, Husam; Voss, Courtney; Chin‐Yee, Benjamin; Stogios, P.J.; Seitova, Almagul; Hutchinson, A.; Yusifov, Farhad; Skarina, T.; Evdokimova, E.; Ackloo, Suzanne; Lowes, Lori E.; Hedley, Benjamin D.; Bhayana, Vipin; Chin‐Yee, Ian; Li, Shawn S.‐C.

doi:10.1016/j.crmeth.2021.100011

Cited by 12 publications

(19 citation statements)

References 27 publications

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“…Importantly, none of the healthy serum samples generated scores >0 for either PS-spike or PS-control microbeads (Figure D), whereas all of the COVID-19 positive samples registered scores of >1 for PS-spike beads, and generally 0 for PS-control microbeads. When titrating positive serum with healthy serum, we found that CAT scores remained positive down to ∼1:10 dilutions (Figure S7), consistent with recent studies reporting traditional well-plate format agglutination assays for SARS-CoV-2 antibodies. , However, given that every sample or analytical standard would have a unique antibody composition (i.e., unique IgG:IgM:IgA:IgD:IgE ratio and total Ig/mL, both of which vary over time as the immune response evolves), it is not feasible or relevant to definitively report assay detection or quantitation limits, consistent with other reports. , For several samples, the PS-control beads registered a score of 1, and these samples each required multiple rounds of centrifugation to clarify precipitate materials prior to testing. Similarly, we also tested some additional negative samples prepared as plasma which showed nonspecific positive signals on both PS-Spike and PS-Control microbeads (Figure S8); for this reason, the current assay format is only appropriate for testing with fasting serum samples, similar for a range of standard blood tests .…”

Section: Resultssupporting

confidence: 82%

Column Agglutination Assay Using Polystyrene Microbeads for Rapid Detection of Antibodies against SARS-CoV-2

Kesarwani

Walker

Henderson

et al. 2022

ACS Appl. Mater. Interfaces

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Rapid serology platforms are essential in disease pandemics for a variety of applications, including epidemiological surveillance, contact tracing, vaccination monitoring, and primary diagnosis in resource-limited areas. Laboratory-based enzymelinked immunosorbent assay (ELISA) platforms are inherently multistep processes that require trained personnel and are of relatively limited throughput. As an alternative, agglutination-based systems have been developed; however, they rely on donor red blood cells and are not yet available for high-throughput screening. Column agglutination tests are a mainstay of pretransfusion blood typing and can be performed at a range of scales, ranging from manual through to fully automated testing. Here, we describe a column agglutination test using colored microbeads coated with recombinant SARS-CoV-2 spike protein that agglutinates when incubated with serum samples collected from patients recently infected with SARS-CoV-2. After confirming specific agglutination, we optimized centrifugal force and time to distinguish samples from uninfected vs SARS-CoV-2-infected individuals and then showed concordant results against ELISA for 22 clinical samples, and also a set of serial bleeds from one donor at days 6−10 postinfection. Our study demonstrates the use of a simple, scalable, and rapid diagnostic platform that can be tailored to detect antibodies raised against SARS-CoV-2 and can be easily integrated with established laboratory frameworks worldwide.

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Section: Resultssupporting

confidence: 82%

Column Agglutination Assay Using Polystyrene Microbeads for Rapid Detection of Antibodies against SARS-CoV-2

Kesarwani

Walker

Henderson

et al. 2022

ACS Appl. Mater. Interfaces

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“…First, we aimed to establish a PA test as a virus titration method using hACE2- bound latex beads (hACE2-beads) as a surrogate for the blood cells used in the HA test of influenza viruses. The hACE2-beads were prepared based on the method used for SARS-CoV-2 antigen-coated latex beads 10 (see the Online Methods). A suspension of the prepared beads showed a clear sedimentation pattern after overnight settling at a final concentration of 0.03% (Fig.…”

Section: Establishment Of a Particle Agglutination (Pa) Testmentioning

confidence: 99%

Use of the particle agglutination/particle agglutination-inhibition test for antigenic analysis of SARS-CoV-2

Kobayashi

Matsuyama

Saijo

et al. 2022

Preprint

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The antigenicity of SARS-CoV-2 is a critical issue for the effectiveness of the vaccine, and thus it should be phenotypically evaluated by serological assays as new field isolates emerge. The hemagglutination/hemagglutination-inhibition (HA/HI) tests are well-known as a representative method for antigenic analysis of influenza viruses, but SARS-CoV-2 is unlikely to agglutinate to human or guinea pig red blood cells. Therefore, the antigenic analysis requires complicated enzyme-linked immunosorbent assay (ELISA) or cell-based assays such as the microneutralization assay. In this study, we developed the particle agglutination/particle agglutination-inhibition (PA/PAI) test to easily and rapidly quantify the virus and antibody using human angiotensin-converting enzyme 2 (hACE2)-bound latex beads. The PA titer was positively correlated with the plaque-forming units. The PAI titer using post-infection Syrian hamster antisera clearly revealed the antigenic difference between the omicron and previous variants. The results show the PAI test is useful for easy and rapid antigenic analysis of SARS-CoV-2.

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“…Immunoassays are widely used in biology and medicine for identifying antibodies in samples with common assays, including latex agglutination tests, lateral flow assays, and Luminex bead-based assays. Latex agglutination tests, for example, have been used to reveal the presence of antibodies within samples by the clumping or aggregation of antigen-coated latex beads, and depending on the format of the assay, have been used for determining immunity to viruses including SARS-CoV-2 [27][28][29][30], as well as detection of infection by bacteria or parasites [31,32]. Applying this principle, we reveal a strategy for the generation of epitope displaying polydiacetylene vesicles capable of being used in the visual detection of the presence of specific antibodies by target-specific binding to induce aggregation.…”

Section: Introductionmentioning

confidence: 99%

Peptide Linked Diacetylene Amphiphiles for Detection of Epitope Specific Antibodies

2022

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Antibodies produced in response to adaptive immunity provide a receptor with multiple sites for binding to a distinct epitope of an antigen. Determining antibody levels to specific antigens has important clinical applications in assessing immune status or deficiency, monitoring infectious or autoimmune diseases, and diagnosing allergies. Leveraging that a specific antibody will bind to a distinct small peptide epitope without requiring the entire antigen to be present, we demonstrate in this work a proof-of-concept assay to detect the presence of an antibody by using peptide epitopes linked to an amphiphile to generate a vesicle-based sensing system. By affording multiple copies of the epitope site on the vesicle, we revealed that the vesicles visibly aggregate in response to an antibody specific for that epitope due to multivalent binding provided by the antibody. We also uncovered the role of peptide surface density in providing accessible epitopes on the vesicles for antibody binding. In summary, using a peptide derived from the coat protein of human influenza virus directly linked to a diacetylene-containing amphiphile afforded peptide-laden vesicles that proved capable of detecting the presence of antibodies specific for human influenza hemagglutinin.

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Rapid and accurate agglutination-based testing for SARS-CoV-2 antibodies

Cited by 12 publications

References 27 publications

Column Agglutination Assay Using Polystyrene Microbeads for Rapid Detection of Antibodies against SARS-CoV-2

Column Agglutination Assay Using Polystyrene Microbeads for Rapid Detection of Antibodies against SARS-CoV-2

Use of the particle agglutination/particle agglutination-inhibition test for antigenic analysis of SARS-CoV-2

Peptide Linked Diacetylene Amphiphiles for Detection of Epitope Specific Antibodies

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