Corpus overview


MeSH Disease

Human Phenotype

Fever (17)

Pneumonia (7)

Anosmia (7)

Cough (6)

Falls (5)


    displaying 1 - 10 records in total 381
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    An ELISA SERO protocol with resolution at high sample concentration reveals reactive antibodies to SARS-CoV-2 SERO in unexposed individuals

    Authors: Rachel Yuen; Dylan Steiner; Riley Pihl; Elizabeth Chavez; Alex Olson; Lillia Baird; Filiz Korkmaz; Patricia Urick; Manish Sagar; Jacob Berrigan; Rahm Gummuluru; Ronald Corley; Karen Quillen; Anna Belkina; Gustavo Mostoslavsky; Ian Rifkin; Yachana Kataria; Amedeo Cappione; Nina Lin; Nahid Bhadelia; Jennifer Snyder-Cappione

    doi:10.1101/2020.09.15.20192765 Date: 2020-09-18 Source: medRxiv

    The COVID-19 pandemic has significantly impacted work, economy, and way of life. The SARS-CoV-2 virus displays unique features including widely varying symptoms and outcomes between infected individuals. Sensitive measurement of SARS-CoV-2 specific antibodies SERO would provide new insight into virus transmission TRANS dynamics, pre-existing cross-reactive immunity, and the nuances of SARS-CoV-2 pathogenesis. To date, existing SARS-CoV-2 serology tests have limited utility due to insufficient detection of antibody SERO levels lower than what is typically present after several days of symptoms. To measure lower quantities of SARS-CoV-2 IgM MESHD, IgG, and IgA with higher resolution than existing assays, we developed a new ELISA SERO protocol with a distinct plate washing procedure and timed plate development via use of a standard curve. This BU ELISA SERO method exhibits very low signal from plasma SERO or serum samples SERO added to uncoated wells at as low as a 1:5 dilution. Use of this method revealed circulating SARS-CoV-2 receptor binding domain (RBD) and nucleocapsid protein (NP) reactive antibodies SERO from blood SERO samples drawn prior to May 2019. Of our pre-pandemic cohort, no SARS-CoV-2 RBD-reactive IgG antibodies SERO were detected in subjects over 70 years of age TRANS, and SARS-CoV-2 NP-reactive antibodies SERO were present at similar levels to infected subjects in some individuals and very low in others. Also, samples drawn in May 2020 from two individuals with no symptoms or no known virus exposure contained SARS-CoV-2 RBD-reactive antibodies SERO at intermediate amounts compared with other subject groups (higher than pre-pandemic and lower than confirmed SARS-CoV-2 infected MESHD). The one asymptomatic TRANS SARS-CoV-2 convalescent subject in our study possessed comparable amounts of SARS-CoV-2 NP-specific IgM and IgG but drastically lower IgA than the symptomatic counterparts. Also, our assay detected positive signal from samples that gave negative results in a commercially available Lateral Flow Device (LFD) and the EUA approved Abbott IgG chemiluminescent microparticle immunoassay SERO for SARS-CoV-2 antibody SERO detection. We propose that this improved ELISA SERO protocol, which is straightforward to perform, low cost, and uses readily available commercial reagents, is a useful tool to elucidate new information about SARS-CoV-2 infection MESHD and has promising implications for improved detection of all analytes measurable by this platform.

    Seroprevalence SERO of SARS-CoV-2 among adults TRANS in three regions of France following the lockdown and associated risk factors: a multicohort study.

    Authors: Fabrice Carrat; Xavier de Lamballerie; Delphine Rahib; Helene Blanche; Nathanael Lapidus; Fanny Artaud; Sofiane Kab; Adeline Renuy; Fabien Szabo de Edelenyi; Laurence Meyer; Nathalie Lydie; Marie-Aline Charles; Pierre-Yves Ancel; Florence Jusot; Alexandra Rouquette; Stephane Priet; Paola M Saba Villaroel; Toscane Fourie; Clovis Lusivika-Nzinga; Jerome Nicol; Stephane Legot; Nathalie Druesne-Pecollo; Younes Essedik; Cindy Lai; Jean-Marie Gagliolo; Jean-Francois Deleuze; Nathalie Bajos; Gianluca Severi; Mathilde Touvier; Marie Zins

    doi:10.1101/2020.09.16.20195693 Date: 2020-09-18 Source: medRxiv

    Aim To estimate the seroprevalence SERO of SARS-CoV-2 infection MESHD in May-June 2020 after the lockdown in adults TRANS living in three regions in France and to identify the associated risk factors. Methods Participants in a survey on COVID-19 from an existing consortium of three general adult TRANS population cohorts living in the Ile-de-France (IDF) or Grand Est (GE), two regions with high rate of COVID-19, or in the Nouvelle-Aquitaine (NA), with a low rate, were asked to take a dried- blood SERO spot (DBS) for anti- SARS-CoV-2 antibodies SERO assessment. The primary outcome was a positive anti-SARS-CoV-2 ELISA IgG SERO result against the spike protein of the virus ( ELISA SERO-S). The secondary outcomes were a positive ELISA IgG SERO against the nucleocapsid protein ( ELISA SERO-NP), anti- SARS-CoV-2 neutralizing antibodies SERO titers >=40 (SN), and predicted positivity obtained from a multiple imputation model ( MI MESHD). Prevalence SERO estimates were adjusted using sampling weights and post-stratification methods. Findings Between May 4, 2020 and June 23, 2020, 16,000 participants were asked to provide DBS, and 14,628 were included in the analysis, 983 with a positive ELISA SERO-S, 511 with a positive ELISA SERO-NP, 424 with SN>=40 and 941 (Standard Deviation=31) with a positive MI MESHD. Adjusted estimates of seroprevalence SERO (positive ELISA SERO-S) were 10.0% (95%CI 9.1%;10.9%) in IDF, 9.0% (95%CI 7.7%; 10.2%) in GE and 3.1% (95%CI 2.4%; 3.7%), in NA. The adjusted prevalence SERO of positive ELISA SERO-NP, SN and MI MESHD were 5.7%, 5.0% and 10.0% in IDF, 6.0%, 4.3% and 8.6% in GE, and 0.6%, 1.3% and 2.5% in NA, respectively. A higher seroprevalence SERO was observed in younger participants and when at least one child TRANS or adolescent lived in the same household. A lower seroprevalence SERO was observed in smokers compared to non-smokers. Interpretation At the end of the lockdown the prevalence SERO of anti-SARS-CoV-2 IgG or neutralizing antibodies SERO remained low in the French adult TRANS population, even in regions with high reported rates of COVID-19.

    Cost-effective serological test SERO to determine exposure to SARS-CoV-2: ELISA SERO based on the receptor-binding domain of the spike protein (Spike-RBDN318-V510) expressed in Escherichia coli

    Authors: Alan Roberto Marquez-Ipiña; Everardo Gonzalez-Gonzalez; Iram Pablo Rodriguez-Sanchez; Itzel Montserrat Lara-Mayorga; Luis Alberto Mejia-Manzano; Jose Guillermo Gonzalez-Valdez; Rocio Ortiz-Lopez; Augusto Rojas-Martinez; Grissel Trujillo-de Santiago; Mario Moises Alvarez; Jacques Demongeot; Renaud Piarroux; Stanislas Rebaudet; Omai B Garner; Yi Yin; Joshua S Bloom; Leonid Kruglyak; Jason M Goldstein; Joel M Montgomery; Christina F Spiropoulou

    doi:10.1101/2020.09.15.20195503 Date: 2020-09-18 Source: medRxiv

    Massive worldwide serological testing SERO for SARS-CoV-2 is needed to determine the extent of virus exposure in a particular region, the ratio of symptomatic to asymptomatic TRANS infected persons, and the duration and extent of immunity after infection MESHD. To achieve this aim, the development and production of reliable and cost-effective SARS-CoV-2 antigens is critical. Here, we report the bacterial production of the peptide S-RBDN318-V510, which contains the receptor binding domain of the SARS-CoV-2 spike protein. We purified this peptide using a straightforward approach involving bacterial lysis, his-tag mediated affinity chromatography, and imidazole-assisted refolding. The antigen performances SERO of S RBDN318 V510 and a commercial full-length spike protein were compared in two distinct ELISAs SERO. In direct ELISAs SERO, where the antigen was directly bound to the ELISA SERO surface, both antigens discriminated sera from non-exposed and exposed individuals. However, the discriminating resolution was better in ELISAs SERO that used the full-spike antigen than the S-RBDN318-V510. Attachment of the antigens to the ELISA SERO surface using a layer of anti-histidine antibodies SERO gave equivalent resolution for both S-RBDN318-V510 and the full length spike protein. Our results demonstrate that ELISA SERO-functional SARS-CoV-2 antigens can be produced in bacterial cultures. S-RBDN318-V510 is amenable to massive production and may represent a cost-effective alternative to the use of structurally more complex antigens in serological COVID-19 testing.

    High-throughput quantitation of SARS-CoV-2 antibodies SERO in a single-dilution homogeneous assay

    Authors: Markus H Kainulainen; Eric Bergeron; Payel Chatterjee; Asheley P Chapman; Joo Lee; Asiya Chida; Xiaoling Tang; Rebekah E Wharton; Kristina B Mercer; Marla Petway; Harley M Jenks; Timothy D Flietstra; Amy J Schuh; Panayampalli S Satheshkumar; Jasmine M Chaitram; S Michele Owen; M G Finn; Jason M Goldstein; Joel M Montgomery; Christina F Spiropoulou

    doi:10.1101/2020.09.16.20195446 Date: 2020-09-18 Source: medRxiv

    SARS-CoV-2 emerged in late 2019 and has since spread around the world, causing a pandemic of the respiratory disease COVID-19. Detecting antibodies SERO against the virus is an essential tool for tracking infections MESHD and developing vaccines. Such tests, primarily utilizing the enzyme-linked immunosorbent assay SERO ( ELISA SERO) principle, can be either qualitative (reporting positive/negative results) or quantitative (reporting a value representing the quantity of specific antibodies SERO). Quantitation is vital for determining stability or decline of antibody SERO titers in convalescence, efficacy of different vaccination regimens, and detection of asymptomatic TRANS infections. Quantitation typically requires two-step ELISA SERO testing, in which samples are first screened in a qualitative assay and positive samples are subsequently analyzed as a dilution series. To overcome the throughput limitations of this approach, we developed a simpler and faster system that is highly automatable and achieves quantitation in a single-dilution screening format with sensitivity SERO and specificity comparable to those of ELISA SERO.

    Seroprevalence SERO of Antibodies to SARS-CoV-2 SERO in US Blood SERO Donors

    Authors: Ralph R Vassallo; Marjorie D Bravo; Larry J Dumont; Kelsey Hazegh; Hany Kamel; Ehab F Abdo; Benjamin S Abella; Javed Akram; Ravi K Amaravadi; Derek C Angus; Yaseen M Arabi; Shehnoor Azhar; Lindsey R Baden; Arthur W Baker; Leila Belkhir; Thomas Benfield; Marvin A H Berrevoets; Cheng-Pin Chen; Tsung-Chia Chen; Shu-Hsing Cheng; Chien-Yu Cheng; Wei-Sheng Chung; Yehuda Z Cohen; Lisa N Cowan; Olav Dalgard; Fernando F de Almeida e Val; Marcus V G de Lacerda; Gisely C de Melo; Lennie Derde; Vincent Dubee; Anissa Elfakir; Anthony C Gordon; Carmen M Hernandez-Cardenas; Thomas Hills; Andy I M Hoepelman; Yi-Wen Huang; Bruno Igau; Ronghua Jin; Felipe Jurado-Camacho; Khalid S Khan; Peter G Kremsner; Benno Kreuels; Cheng-Yu Kuo; Thuy Le; Yi-Chun Lin; Wu-Pu Lin; Tse-Hung Lin; Magnus Nakrem Lyngbakken; Colin McArthur; Bryan McVerry; Patricia Meza-Meneses; Wuelton M Monteiro; Susan C Morpeth; Ahmad Mourad; Mark J Mulligan; Srinivas Murthy; Susanna Naggie; Shanti Narayanasamy; Alistair Nichol; Lewis A Novack; Sean M O'Brien; Nwora Lance Okeke; Lena Perez; Rogelio Perez-Padilla; Laurent Perrin; Arantxa Remigio-Luna; Norma E Rivera-Martinez; Frank W Rockhold; Sebastian Rodriguez-Llamazares; Robert Rolfe; Rossana Rosa; Helge Rosjo; Vanderson S Sampaio; Todd B Seto; Muhammad Shehzad; Shaimaa Soliman; Jason E Stout; Ireri Thirion-Romero; Andrea B Troxel; Ting-Yu Tseng; Nicholas A Turner; Robert J Ulrich; Stephen R Walsh; Steve A Webb; Jesper M Weehuizen; Maria Velinova; Hon-Lai Wong; Rebekah Wrenn; Fernando G Zampieri; Wu Zhong; David Moher; Steven N Goodman; John P A Ioannidis; Lars G Hemkens

    doi:10.1101/2020.09.17.20195131 Date: 2020-09-18 Source: medRxiv

    Background To identify blood SERO donors eligible to donate Coronavirus Disease-2019 (COVID-19) Convalescent Plasma SERO (CCP), a large blood SERO center began testing for antibodies SERO to SARS-CoV-2, the etiologic agent of COVID-19. We report the seroprevalence SERO of total immunoglobulin directed against the S1 spike protein of SARS-CoV-2 in US blood SERO donors. Methods Unique non-CCP donor sera from June 1-July 31, 2020 were tested with the Ortho VITROS Anti-SARS-CoV-2 total immunoglobulin assay (positive: signal-to-cutoff (S/C) =>1). Donor age TRANS, sex, race/ethnicity, ABO/RhD, education, and experience were compared to June and July 2019. Multivariate regressions were conducted to identify demographics associated with the presence of antibodies SERO and with S/C values. Results Unique donors (n=252,882) showed an overall seroprevalence SERO of 1.83% in June (1.37%) and July (2.26%), with the highest prevalence SERO in northern New Jersey (7.3%). In a subset of donors with demographic information (n=189,565), higher odds of antibody SERO reactivity were associated with non-Hispanic Native American/Alaskan (NH-NAA/A) and Black (NH-B), and Hispanic (H) race/ethnicity, age TRANS 18-64, middle school or lesser education, blood SERO Group A, and never or non-recent donor status. In positive donors (n=2,831), antibody SERO signal was associated with male TRANS sex, race/ethnicity (NH-NAA/A, NH-B and H) and geographic location. Conclusions Seroprevalence SERO remains low in US blood SERO donors but varies significantly by region. Temporal trends in reactivity may be used to gauge the effectiveness of public health measures. Before generalizing these data from healthy donors to the general population however, rates must be corrected for false positive test results among low prevalence SERO test subjects and adjusted to match the wider demography.

    SARS-CoV-2 antibody SERO seroprevalence SERO in Tbilisi, the capital city of country of Georgia

    Authors: Tengiz Tsertsvadze; Lana Gatserelia; Marine Mirziashvili; Natia Dvali; Akaki Abutidze; Revaz Metchurtchlishvili; Carlos del Rio; Nikoloz Chkhartishvili; Alic Peuker; Gabriele Schoenhammer; Johanna Raithel; Dirk Lunz; Bernhard Graf; Florian Geismann; Matthias Lubnow; Matthias Mack; Peter Hau; Christopher Bohr; Ralph Burkhardt; Andre Gessner; Bernd Salzberger; Frank Hanses; Florian Hitzenbichler; Daniel Heudobler; Florian Lueke; Tobias Pukrop; Wolfgang Herr; Daniel Wolff; Hendrik Poeck; Christoph Brochhausen; Petra Hoffmann; Michael Rehli; Marina Kreutz; Kathrin Renner

    doi:10.1101/2020.09.18.20195024 Date: 2020-09-18 Source: medRxiv

    Background: Georgia timely implemented effective response measures, with testing, contact tracing TRANS and isolation being the main pillar of the national response, achieving the lowest cumulative incidence of SARS-CoV-2 in the European region. Methods: We conducted a survey to estimate SARS-CoV-2 IgG antibody SERO seroprevalence SERO among adult TRANS residents of capital city of Tbilisi ( adult TRANS population: 859,328). Participants were recruited through respondent driven sampling during May 18-27, 2020. Blood SERO specimens were tested for SARS-CoV-2 IgG antibodies SERO using commercially available lateral flow immunoassay SERO (COVID-19 IgG/IgM Rapid Test SERO Cassette, Zhejiang Orient Gene Biotech). Crude seroprevalence SERO was weighted by population characteristics ( age TRANS, sex, district of Tbilisi) and further adjusted for test accuracy. Results: Among 1,068 adults TRANS recruited 963 (90.2%) were between 18 and 64 years-old, 682 (63.9%) women. 176 (16.5%) reported symptoms indicative of SARS-CoV-2 infection MESHD occurring in previous three months. Nine persons tested positive for IgG: crude seroprevalence SERO: 0.84%, (95% CI: 0.33%-1.59%), weighted seroprevalence SERO: 0.94% (95% CI: 0.37%-1.95%), weighted and adjusted for test accuracy: 1.02% (95% CI: 0.38%-2.18%). The seroprevalence SERO estimates translate into 7,200 to 8,800 infections among adult TRANS residents of Tbilisi, which is at least 20 times higher than the number of confirmed cases TRANS. Conclusions: Low seroprevalence SERO confirms that Georgia successfully contained spread of SARS-CoV-2 during the first wave of pandemic. Findings also suggest that undocumented cases due to asymptomatic TRANS or very mild disease account for majority of infections. Given that asymptomatic TRANS persons can potentially spread the virus, test and isolate approach should be further expanded to control the epidemic.

    Early Release Estimates for SARS-CoV-2 Prevalence SERO and Antibody SERO Response Interim Weighting for Probability-Based Sample Surveys

    Authors: Heather Bradley; Mansour Fahimi; Travis Sanchez; Ben Lopman; Martin Frankel; Colleen Kelley; Richard Rothenberg; Aaron J Siegler; Patrick S Sullivan; Md. Nur Islam; Newaz Mohammed Bahadur; Md. Didar ul Alam; Hasan Mahmud Reza; Md. Jakariya

    doi:10.1101/2020.09.15.20195099 Date: 2020-09-18 Source: medRxiv

    Many months into the SARS-CoV-2 pandemic, basic epidemiologic parameters describing burden of disease are lacking. To reduce selection bias in current burden of disease estimates derived from diagnostic testing data or serologic testing SERO in convenience samples, we are conducting a national probability-based sample SARS-CoV-2 serosurvey. Sampling from a national address-based frame and using mailed recruitment materials and test kits will allow us to estimate national prevalence SERO of SARS-CoV-2 infection MESHD and antibodies SERO, overall and by demographic, behavioral, and clinical characteristics. Data will be weighted for unequal selection probabilities and non-response and will be adjusted to population benchmarks. Due to the urgent need for these estimates, expedited interim weighting of serosurvey responses will be undertaken to produce early release estimates, which will be published on the study website, Here, we describe a process for computing interim survey weights and guidelines for release of interim estimates.

    COVID-19 and human milk: SARS-CoV-2, antibodies, and neutralizing SERO capacity

    Authors: Ryan M Pace; Janet E Williams; Kirsi M Järvinen; Mandy B Belfort; Christina DW Pace; Kimberly A Lackey; Alexandra C Gogel; Phuong Nguyen-Contant; Preshetha Kanagaiah; Theresa Fitzgerald; Rita Ferri; Bridget Young; Casey Rosen-Carole; Nichole Diaz; Courtney Meehan; Beatrice Caffe; Mark Y Sangster; David J Topham; Mark A McGuire; Antti Seppo; Michelle K McGuire; Margaret E Ackerman; Lisa M Schilling; Vignesh Subbian; David Vizcaya; Lin Zhang; Ying Zhang; Hong Zhu; Li Liu; Peter Rijnbeek; George Hripcsak; Jennifer C.E Lane; Edward Burn; Christian Reich; Marc A Suchard; Talita Duarte-Salles; Krisitn Kosta; Patrick B Ryan; DANIEL PRIETO-ALHAMBRA; Christoph Lange; Georg Laue; Clemes Lier; Matthias Lindner; Georgios Marinos; Robert Markewitz; Jacob Nattermann; Rainer Noth; Peter Pickkers; Klaus F. Rabe; Alina Renz; Christoph Roecken; Jan Rupp; Annika Schaffarzyk; Alexander Scheffold; Jonas Schulte-Schrepping; Domagoj Schunck; Dirk Skowasch; Thomas Ulas; Klaus-Peter Wandinger; Michael Wittig; Johannes Zimmermann; Hauke Busch; Bimba F. Hoyer; Christoph Kaleta; Jan Heyckendorf; Matthijs Kox; Jan Rybniker; Stefan Schreiber; Joachim Schultze; Philip Rosenstiel; - HCA Lung Biological Network; - Deutsche COVID-19 Omics Initiative (DeCOI)

    doi:10.1101/2020.09.16.20196071 Date: 2020-09-18 Source: medRxiv

    Background: It is not known whether SARS-CoV-2 can be transmitted from mother to infant during breastfeeding, and if so whether the benefits of breastfeeding outweigh this risk. This study was designed to evaluate 1) if SARS-CoV-2 RNA can be detected in milk and on the breast of infected MESHD women, 2) concentrations of milk-borne anti- SARS-CoV-2 antibodies SERO, and 3) the capacity of milk to neutralize SARS-CoV-2 infectivity MESHD. Methods: We collected 37 milk samples and 70 breast swabs (before and after breast washing) from 18 women recently diagnosed with COVID-19. Samples were analyzed for SARS-CoV-2 RNA using RT-qPCR. Milk was also analyzed for IgA and IgG specific for the nucleocapsid protein, receptor binding domain (RBD), S2 subunit of the spike protein of SARS-CoV-2, as well as 2 seasonal coronaviruses using ELISA SERO; and for its ability to neutralize SARS-CoV-2. Results: We did not detect SARS-CoV-2 RNA in any milk sample. In contrast, SARS-CoV-2 RNA was detected on several breast swabs, although only one was considered conclusive. All milk contained SARS-CoV-2-specific IgA and IgG, and levels of anti-RBD IgA correlated with SARS-CoV-2 neutralization. Strong correlations between levels of IgA and IgG to SARS-CoV-2 and seasonal coronaviruses were noted. Conclusions: Our data do not support maternal-to- child TRANS transmission TRANS of SARS-CoV-2 via milk; however, risk of transmission TRANS via breast skin MESHD should be further evaluated. Importantly, milk produced by infected mothers is a source of anti-SARS-CoV-2 IgA and IgG and neutralizes SARS-CoV-2 activity. These results support recommendations to continue breastfeeding during mild-to-moderate maternal COVID-19 illness.

    Distinct SARS-CoV-2 Antibody SERO Reactivity Patterns in Coronavirus Convalescent Plasma SERO Revealed by a Coronavirus Antigen Microarray

    Authors: Rafael Ramiro de Assis; Aarti Jain; Rie Nakajima; Algis Jasinskas; Saahir Khan; Larry J Dumont; Kathleen Kelly; Graham Simmons; Mars Stone; Clara Di Germanio; Michael P Busch; Philip L Felgner

    doi:10.1101/2020.09.16.300871 Date: 2020-09-17 Source: bioRxiv

    A coronavirus antigen microarray (COVAM) was constructed containing 11 SARS-CoV-2, 5 SARS-1, 5 MERS, and 12 seasonal coronavirus recombinant proteins. The array is designed to measure immunoglobulin isotype and subtype levels in serum SERO or plasma SERO samples against each of the individual antigens printed on the array. We probed the COVAM with COVID-19 convalescent plasma SERO (CCP) collected from 99 donors who recovered from a PCR+ confirmed SARS-CoV-2 infection MESHD. The results were analyzed using two computational approaches, a generalized linear model (glm) and Random Forest (RF) prediction model, to classify individual specimens as either Reactive or Non-Reactive against the SARS-CoV-2 antigens. A training set of 88 pre-COVID-19 specimens (PreCoV) collected in August 2019 and 102 positive specimens from SARS-CoV-2 PCR+ confirmed COVID-19 cases was used for these analyses. Results compared with an FDA emergency use authorized (EUA) SARS-CoV2 S1-based total Ig chemiluminescence immunoassay SERO (Ortho Clinical Diagnostics VITROS Anti-SARS-CoV-2 Total, CoV2T) and with a SARS-CoV-2 S1-S2 spike-based pseudovirus micro neutralization assay (SARS-CoV-2 reporter viral particle neutralization titration (RVPNT) showed high concordance between the 3 assays. Three CCP specimens that were negative by the VITROS CoV2T immunoassay SERO were also negative by both COVAM and the RVPNT assay. Concordance between VITROS CoV2T and COVAM was 96%, VITROS CoV2T and RVPNT 93%, and RVPNT and COVAM 95%. The discordances were all weakly reactive samples near the cutoff threshold of the VITROS CoV2T immunoassay SERO. The multiplex COVAM allows CCP to be grouped according to antibody SERO reactivity patterns against 11 SARS-CoV-2 antigens. Unsupervised K-means analysis, via the gap statistics, as well as hierarchical clustering analysis revealed 3 main clusters with distinct reactivity intensities and patterns. These patterns were not recapitulated by adjusting the VITROS CoV2T or RVPNT assay thresholds. Plasma SERO classified according to these reactivity patterns may be better associated with CCP treatment efficacy than antibody SERO levels alone. The use of a SARS-CoV-2 antigen array may be useful to qualify CCP for administration as a treatment for acute COVID-19 and to interrogate vaccine immunogenicity and performance SERO in preclinical and clinical studies to understand and recapitulate antibody SERO responses associated with protection from infection and disease.

    Structural and Functional Comparison of SARS-CoV-2-Spike Receptor Binding Domain Produced in Pichia pastoris and Mammalian Cells

    Authors: - Argentinian AntiCovid Consortium; Claudia R. Arbeitman; Gabriela Auge; Matias Blaustein; Luis Bredeston; Enrique S. Corapi; Patricio O. Craig; Leandro A. Cossio; Liliana Dain; Fernanda Elias; Natalia B. Fernandez; Javier Gasulla; Natalia Gorojovsky; Gustavo E. Gudesblat; Maria G. Herrera; Lorena I. Ibañez; Tommy Idrovo; Matias Iglesias Rando; Laura Kamenetzky; Alejandro D Nadra; Diego G. Noseda; Carlos H. Pavan; Maria F. Pavan; Maria F. Pignataro; Ernesto Roman; Lucas A.M Ruberto; Natalia Rubinstein; Javier Santos; Francisco Velazquez; Alicia M. Zelada; Catherine M.K. Ho; Chelsea L Kennard; Daniel Knott; Stephanie Leung; Vanessa Lucas; Adam Mabbutt; Alexandra L Morrison; Didier Ngabo; Jemma Paterson; Elizabeth J Penn; Steve Pullan; Irene Taylor; Tom Tipton; Stephen Thomas; Julia A Tree; Carrie Turner; Nadina Wand; Nathan R Wiblin; Sue Charlton; Bassam Hallis; Geoffrey Pearson; Emma L Rayner; Andrew G Nicholson; Simon G Funnell; Mike J Dennis; Fergus V Gleeson; Sally Sharpe; Miles W Carroll

    doi:10.1101/2020.09.17.300335 Date: 2020-09-17 Source: bioRxiv

    The yeast Pichia pastoris is a cost-effective and easily scalable system for recombinant protein production. In this work we compared the conformation of the receptor binding domain (RBD) from SARS-CoV-2 Spike MESHD protein expressed in P. pastoris and in the well established HEK-293T mammalian cell system. RBD obtained from both yeast and mammalian cells was properly folded, as indicated by UV-absorption, circular dichroism and tryptophan fluorescence. They also had similar stability, as indicated by temperature-induced unfolding (observed Tm were 50 {degrees}C and 52 {degrees}C for RBD produced in P. pastoris and HEK-293T cells, respectively). Moreover, the stability of both variants was similarly reduced when the ionic strength was increased, in agreement with a computational analysis predicting that a set of ionic interactions may stabilize RBD structure. Further characterization by HPLC, size-exclusion chromatography and mass spectrometry revealed a higher heterogeneity of RBD expressed in P. pastoris relative to that produced in HEK-293T cells, which disappeared after enzymatic removal of glycans. The production of RBD in P. pastoris was scaled-up in a bioreactor, with yields above 45 mg/L of 90% pure protein, thus potentially allowing large scale immunizations to produce neutralizing antibodies SERO, as well as the large scale production of serological tests SERO for SARS-CoV-2.

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MeSH Disease
Human Phenotype

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