A universal method for the functionalization of dyed magnetic microspheres with peptides

Coppock, Matthew B.; Stratis‐Cullum, Dimitra N.

doi:10.1016/j.ymeth.2019.01.014

Cited by 8 publications

(4 citation statements)

References 7 publications

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“…We chose 12 SARS-CoV-2 peptides predicted by VirScan data and the machine learning model to be highly indicative of SARS-CoV-2 exposure history (table S9). These SARS-CoV-2 peptides, two positive control peptides from rhinovirus A and Epstein-Barr virus (EBV) that are recognized in >80% of seropositive individuals by VirScan (7), and a negative control peptide from HIV-1 were coupled to Luminex beads (13). We tested 163 COVID-19 patient samples and 165 pre-COVID-19 era controls for IgG reactivity to the Luminex panel.…”

Section: Machine Learning Guides the Design Of A Luminex Assay For Ra...mentioning

confidence: 99%

“…Multiplexed SARS-CoV-2 peptide epitope assays were built using the peptides listed in table S9. Peptides were synthesized by the Ragon/ MGH Peptide Core Facility with a Proparglyglycine (Pra, X) moiety in the N terminus to facilitate cross-linking to Luminex beads using a "click" chemistry strategy as described previously (13). In brief, Luminex beads were first functionalized with amine-PEG4-azide and then reacted with the peptides to generate 20 different Luminex beads with attached peptides.…”

Section: Luminex Multiplex Peptide Epitope Serology Assaysmentioning

confidence: 99%

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Viral epitope profiling of COVID-19 patients reveals cross-reactivity and correlates of severity

Shrock

Fujimura

Kula

et al. 2020

Science

610

774

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Understanding humoral responses to SARS-CoV-2 is critical for improving diagnostics, therapeutics, and vaccines. Deep serological profiling of 232 COVID-19 patients and 190 pre-COVID-19 era controls using VirScan revealed over 800 epitopes in the SARS-CoV-2 proteome, including 10 epitopes likely recognized by neutralizing antibodies. Pre-existing antibodies in controls recognized SARS-CoV-2 ORF1, while only COVID-19 patients primarily recognized spike and nucleoprotein. A machine learning model trained on VirScan data predicted SARS-CoV-2 exposure history with 99% sensitivity and 98% specificity; a rapid Luminex-based diagnostic was developed from the most discriminatory SARS-CoV-2 peptides. Individuals with more severe COVID-19 exhibited stronger and broader SARS-CoV-2 responses, weaker antibody responses to prior infections, and higher incidence of CMV and HSV-1, possibly influenced by demographic covariates. Among hospitalized patients, males make greater SARS-CoV-2 antibody responses than females.

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Section: Machine Learning Guides the Design Of A Luminex Assay For Ra...mentioning

confidence: 99%

Section: Luminex Multiplex Peptide Epitope Serology Assaysmentioning

confidence: 99%

Viral epitope profiling of COVID-19 patients reveals cross-reactivity and correlates of severity

Shrock

Fujimura

Kula

et al. 2020

Science

610

774

View full text Add to dashboard Cite

show abstract

“…This protocol describes an easily adaptable serological assay to measure antibody levels to multiple antigens simultaneously. The antigen panel we describe here covalently couples recombinant proteins to the magnetic beads, but slightly modified protocols allow for other antigens such as peptides, to also be coupled to the beads [20].…”

Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 Multi-Antigen Serology Assay

Mazhari

Ruybal‐Pesántez

Angrisano

et al. 2021

MPs

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Serology tests are extremely useful for assessing whether a person has been infected with a pathogen. Since the onset of the COVID-19 pandemic, measurement of anti-SARS-CoV-2-specific antibodies has been considered an essential tool in identifying seropositive individuals and thereby understanding the extent of transmission in communities. The Luminex system is a bead-based technology that has the capacity to assess multiple antigens simultaneously using very low sample volumes and is ideal for high-throughput studies. We have adapted this technology to develop a COVID-19 multi-antigen serological assay. This protocol described here carefully outlines recommended steps to optimize and establish this method for COVID-19-specific antibody measurement in plasma and in saliva. However, the protocol can easily be customized and thus the assay is broadly applicable to measure antibodies to other pathogens.

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“…Luminex bead regions were modified with NH2 -PEGx -Azide (MW 1000) and subsequent peptide attachment was conducted by "click" chemistry according to a previously reported protocol [11]. These bead regions (2500 beads/region) were then incubated with varying concentrations of biotinylated protein targets of interest in Protein Free T20 buffer (96-Well plate) at room temperature for two hours followed by washing (2x) with 1:10 diluted Protein Free T20 buffer (diluted with PBS).…”

Section: Luminex Selectivity Assaysmentioning

confidence: 99%

Development and integration of protein catalyzed capture agents as novel receptors for pathogen detection

Winton

Hajian²,

Liu

et al. 2023

Smart Biomedical and Physiological Sensor Technology XX

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For over two years, the world has endured a coronavirus pandemic resulting in over 470 million illnesses, 6 million deaths, and substantial supply chain disruptions. Prior to effective treatments, experts estimated that in the United States alone as many as 20 -30 million tests should be conducted weekly to safely reopen, while highly effective vaccines took nearly a year to enter general distribution. This global event has highlighted the importance of developing and deploying rapid testing and treatment options for emerging pathogens. Currently, antibodies are the gold standard for biorecognition elements used in biosensors and may also be utilized to treat infection. Unfortunately, they can be challenging to mass produce and are sensitive to biological and temperature degradation, limiting broader distribution and equitable global access.Protein Catalyzed Capture agents (PCCs) present an alluring alternative as these small peptide macrocycles are comprised of unnatural amino acids and exhibit thermal and enzymatic stability. Their chemical synthesis enables scalable and reproducible production at diminished cost, while modular functionalities allow versatile applications in sensing and therapeutics, expanding the potential for more ubiquitous access to sensing & treatment options. This work sought to derive PCC receptors with high affinity and specificity for the SARS-CoV-2 spike protein from a one bead one compound peptide library. Chemically synthesized epitope fragments of the SARS-COV-2 spike protein were screened against the library and PCC receptor leads identified through entropically favored, un-catalyzed "click" reactions. Mass spectrometry and multiplex affinity assays then enabled down-selection of promising receptors. Modular chemical modifications were made to selected receptors to enable effective integration into a graphene field effect transistor (gFET) sensor which demonstrated a practical limit of detection of 103 pfu/mL inactivated SARS-CoV-2 virus and the ability to discriminate between SARS-COV-2 and Influenza or Rhino viruses.

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A universal method for the functionalization of dyed magnetic microspheres with peptides

Cited by 8 publications

References 7 publications

Viral epitope profiling of COVID-19 patients reveals cross-reactivity and correlates of severity

Viral epitope profiling of COVID-19 patients reveals cross-reactivity and correlates of severity

SARS-CoV-2 Multi-Antigen Serology Assay

Development and integration of protein catalyzed capture agents as novel receptors for pathogen detection

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