A high-throughput multiplexed microfluidic device for COVID-19 serology assays

Lab on a Chip ◽  
2021 ◽  
Vol 21 (1) ◽  
pp. 93-104
Author(s):  
Roberto Rodriguez-Moncayo ◽  
Diana F. Cedillo-Alcantar ◽  
Pablo E. Guevara-Pantoja ◽  
Oriana G. Chavez-Pineda ◽  
Jose A. Hernandez-Ortiz ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
High Throughput ◽  
Spike Protein ◽  
Serum Samples

A microfluidic device to quantitate antibody reactivities to SARS-CoV-2 antigens: spike protein, RDB, S1 subunit, and nucleocapsid, from 50 serum samples.

scholarly journals A high-throughput cell- and virus-free assay shows reduced neutralization of SARS-CoV-2 variants by COVID-19 convalescent plasma

2021 ◽  
pp. eabi8452
Author(s):  
Craig Fenwick ◽  
Priscilla Turelli ◽  
Céline Pellaton ◽  
Alex Farina ◽  
Jérémy Campos ◽  
...  
Keyword(s):  
High Throughput ◽  
Neutralizing Antibodies ◽  
Competitive Inhibition ◽  
Natural Infection ◽  
Neutralizing Antibody ◽  
Antibody Responses ◽  
Spike Protein ◽  
Serum Samples ◽  
Live Virus ◽  
Protein Variants

The detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific antibodies in the serum of an individual indicates prior infection or vaccination. However, it provides limited insight into the protective nature of this immune response. Neutralizing antibodies recognizing the viral spike protein are more revealing, yet their measurement traditionally requires virus- and cell-based systems that are costly, time-consuming, inflexible, and potentially biohazardous. Here, we present a cell-free quantitative neutralization assay based on the competitive inhibition of trimeric SARS-CoV-2 spike protein binding to the angiotensin converting enzyme 2 (ACE2) receptor. This high-throughput method matches the performance of the gold standard live virus infection assay, as verified with a panel of 206 seropositive donors with varying degrees of infection severity and virus-specific IgG titers, achieving 96.7% sensitivity and 100% specificity. Furthermore, it allows for the parallel assessment of neutralizing activities against multiple SARS-CoV-2 spike protein variants of concern. We used our assay to profile serum samples from 59 patients hospitalized with coronavirus disease 2019 (COVID-19). We found that, although most sera had high activity against the 2019-nCoV parental spike protein and, to a lesser extent, the α (B.1.1.7) variant, only 58% of serum samples could efficiently neutralize a spike protein derivative containing mutations present in the β (B.1.351) variant. Thus, we have developed an assay that can evaluate effective neutralizing antibody responses to SARS-CoV-2 spike protein variants of concern after natural infection and that can be applied to characterize vaccine-induced antibody responses or to assess the potency of monoclonal antibodies.

scholarly journals Measurement of Carcinoembryonic Antigen in Clinical Serum Samples Using a Centrifugal Microfluidic Device

Micromachines ◽  
2018 ◽  
Vol 9 (9) ◽  
pp. 470 ◽  
Author(s):  
Zhigang Gao ◽  
Zongzheng Chen ◽  
Jiu Deng ◽  
Xiaorui Li ◽  
Yueyang Qu ◽  
...  
Keyword(s):  
Carcinoembryonic Antigen ◽  
Microfluidic Device ◽  
High Throughput ◽  
Classical Method ◽  
Gradient Centrifugation ◽  
Serum Samples ◽  
Clinical Method ◽  
Alternative Approach ◽  
Long Time ◽  
Good Agreement

Carcinoembryonic antigen (CEA) is a broad-spectrum tumor marker used in clinical applications. The primarily clinical method for measuring CEA is based on chemiluminescence in serum during enzyme-linked immunosorbent assays (ELISA) in 96-well plates. However, this multi-step process requires large and expensive instruments, and takes a long time. In this study, a high-throughput centrifugal microfluidic device was developed for detecting CEA in serum without the need for cumbersome washing steps normally used in immunoreactions. This centrifugal microdevice contains 14 identical pencil-like units, and the CEA molecules are separated from the bulk serum for subsequent immunofluorescence detection using density gradient centrifugation in each unit simultaneously. To determine the optimal conditions for CEA detection in serum, the effects of the density of the medium, rotation speed, and spin duration were investigated. The measured values from 34 clinical serum samples using this high-throughput centrifugal microfluidic device showed good agreement with the known values (average relative error = 9.22%). These results indicate that the high-throughput centrifugal microfluidic device could provide an alternative approach for replacing the classical method for CEA detection in clinical serum samples.

scholarly journals Detection of SARS-CoV-2 IgG Targeting Nucleocapsid or Spike Protein by Four High-Throughput Immunoassays Authorized for Emergency Use

2020 ◽  
Vol 58 (11) ◽  
Author(s):  
Harry E. Prince ◽  
Tara S. Givens ◽  
Mary Lapé-Nixon ◽  
Nigel J. Clarke ◽  
Dale A. Schwab ◽  
...  
Keyword(s):  
High Throughput ◽  
Qualitative Agreement ◽  
Nucleocapsid Protein ◽  
Spike Protein ◽  
True Positive ◽  
Serum Samples ◽  
Igg Antibody ◽  
Positive Reactivity ◽  
Very High ◽  
Abbott Architect

ABSTRACT A total of 1,200 serum samples that were tested for SARS-CoV-2 IgG antibody using the Abbott Architect immunoassay targeting the nucleocapsid protein were run in 3 SARS-CoV-2 IgG immunoassays targeting spike proteins (DiaSorin Liaison, Ortho Vitros, and Euroimmun). Consensus-positive and consensus-negative interpretations were defined as qualitative agreement in at least 3 of the 4 assays. Agreement of the 4 individual assays with a consensus-negative interpretation (n = 610) ranged from 96.7% to 100%, and agreement with a consensus-positive interpretation (n = 584) ranged from 94.3% to 100%. Laboratory-developed inhibition assays were utilized to evaluate 49 consensus-negative samples that were positive in only one assay; true-positive reactivity was confirmed in only 2 of these 49 (4%) samples. These findings demonstrate very high levels of agreement among 4 SARS-CoV-2 IgG assays authorized for emergency use, regardless of antigen target or assay format. Although false-positive reactivity was identified, its occurrence was rare (no more than 1.7% of samples for a given assay).

scholarly journals Quantification of a Proteotypic Peptide from Protein C Inhibitor by Liquid Chromatography–Free SISCAPA-MALDI Mass Spectrometry: Application to Identification of Recurrence of Prostate Cancer

2013 ◽  
Vol 59 (10) ◽  
pp. 1514-1522 ◽  
Author(s):  
Morteza Razavi ◽  
Lisa DS Johnson ◽  
Julian J Lum ◽  
Gary Kruppa ◽  
N Leigh Anderson ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Liquid Chromatography ◽  
High Throughput ◽  
Protein C ◽  
Specific Antigen ◽  
Serum Concentrations ◽  
Serum Samples ◽  
Biomarker Validation ◽  
Protein C Inhibitor ◽  
Proteotypic Peptide

BACKGROUND Biomarker validation remains one of the most challenging constraints to the development of new diagnostic assays. To facilitate biomarker validation, we previously developed a chromatography-free stable isotope standards and capture by antipeptide antibodies (SISCAPA)-MALDI assay allowing rapid, high-throughput quantification of protein analytes in large sample sets. Here we applied this assay to the measurement of a surrogate proteotypic peptide from protein C inhibitor (PCI) in sera from patients with prostate cancer. METHODS A 2-plex SISCAPA-MALDI assay for quantification of proteotypic peptides from PCI and soluble transferrin receptor (sTfR) was used to measure these peptides in 159 trypsin-digested sera collected from 51 patients with prostate cancer. These patients had been treated with radiation with or without neoadjuvant androgen deprivation. RESULTS Patients who experienced biochemical recurrence of prostate cancer showed decreased serum concentrations of the PCI peptide analyte within 18 months of treatment. The PCI peptide concentrations remained increased in the sera of patients who did not experience cancer recurrence. Prostate-specific antigen concentrations had no predictive value during the same time period. CONCLUSIONS The high-throughput, liquid chromatography–free SISCAPA-MALDI assay is capable of rapid quantification of proteotypic PCI and sTfR peptide analytes in complex serum samples. Decreased serum concentrations of the PCI peptide were found to be related to recurrence of prostate cancer in patients treated with radiation with or without hormone therapy. However, a larger cohort of patients will be required for unequivocal validation of the PCI peptide as a biomarker for clinical use.

scholarly journals HiSpike Method for High-Throughput Cost Effective Sequencing of the SARS-CoV-2 Spike Gene

2022 ◽  
Vol 8 ◽  
Author(s):  
Ephraim Fass ◽  
Gal Zizelski Valenci ◽  
Mor Rubinstein ◽  
Paul J. Freidlin ◽  
Shira Rosencwaig ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Cost Effective ◽  
Developed Countries ◽  
Spike Protein ◽  
Medical Community ◽  
Effective Vaccine ◽  
Gene Variants ◽  
Spike Gene ◽  
Field Samples

The changing nature of the SARS-CoV-2 pandemic poses unprecedented challenges to the world's health systems. Emerging spike gene variants jeopardize global efforts to produce immunity and reduce morbidity and mortality. These challenges require effective real-time genomic surveillance solutions that the medical community can quickly adopt. The SARS-CoV-2 spike protein mediates host receptor recognition and entry into the cell and is susceptible to generation of variants with increased transmissibility and pathogenicity. The spike protein is the primary target of neutralizing antibodies in COVID-19 patients and the most common antigen for induction of effective vaccine immunity. Tight monitoring of spike protein gene variants is key to mitigating COVID-19 spread and generation of vaccine escape mutants. Currently, SARS-CoV-2 sequencing methods are labor intensive and expensive. When sequence demands are high sequencing resources are quickly exhausted. Consequently, most SARS-CoV-2 strains are sequenced in only a few developed countries and rarely in developing regions. This poses the risk that undetected, dangerous variants will emerge. In this work, we present HiSpike, a method for high-throughput cost effective targeted next generation sequencing of the spike gene. This simple three-step method can be completed in < 30 h, can sequence 10-fold more samples compared to conventional methods and at a fraction of their cost. HiSpike has been validated in Israel, and has identified multiple spike variants from real-time field samples including Alpha, Beta, Delta and the emerging Omicron variants. HiSpike provides affordable sequencing options to help laboratories conserve resources for widespread high-throughput, near real-time monitoring of spike gene variants.

scholarly journals Generation of Gradients on a Microfluidic Device: Toward a High-Throughput Investigation of Spermatozoa Chemotaxis

PLoS ONE ◽  
2015 ◽  
Vol 10 (11) ◽  
pp. e0142555 ◽  
Author(s):  
Yi Zhang ◽  
Rong-Rong Xiao ◽  
Tailang Yin ◽  
Wei Zou ◽  
Yun Tang ◽  
...  
Keyword(s):  
Microfluidic Device ◽  
High Throughput

Abstract 444: Continuous, high throughput microfluidic device to monitor circulating tumor cells in cancer patients

2019 ◽  
Author(s):  
Kaylee Smith ◽  
Tae Hyun Kim ◽  
Costanza Paoletti ◽  
Douglas H. Thamm ◽  
Daniel F. Hayes ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
Microfluidic Device ◽  
High Throughput
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