Scientific poster titled "From HPV to COVID-19 and beyond: Leveraging the power of serology and standards"
Scientific poster presented at AACR23. The below text details the poster's contents.

Background​

  • The SARS-CoV-2 pandemic created a crucial need for high quality serology assays and critical reagents to evaluate seroprevalence and antiviral immune responses ​
  • The initial flood of serology assays entering the market with inadequate performance emphasized the urgent need of standardization and independent evaluation​
Graphic representation of COVID-19 pandemic response workflow

HPV Serology standards expertise was critical for rapid pivot to respond to the pandemic​

  • The HPV Serology Lab (HPVSL) at the Frederick National Laboratory (FNL) for Cancer Research leveraged their experience developing HPV serology standards and partnerships to quickly pivot to COVID-19​
  • In March 2020, the HPV Serology Lab began to develop assays and standards for SARS-CoV-2 serology​
  • In parallel, HPVSL became a critical members of a trans-governmental collaboration established to:​
  • Develop evaluation/validation panels. ​
  • Evaluate commercial SARS-CoV-2 serology assay performance​
  • Inform FDA regulatory decisions for Emergency Use Authorization (EUA)

HPVSL collaborated with NCI to establish the Serological Sciences Network (SeroNet)

SeroNet: The largest US network working on understanding immune response to the virus and vaccines

  • Worked with Capacity Building Centers (CBC) to develop and expand the nation’s capacity for ​
    SARS-CoV-2 serologic testing​

  • Developed the infrastructure to collect and validate patient serology samples for the development of evaluation/validation panels and standards

FNL Serology Laboratory​

  • Sample/assay validation and serology panel construction​

  • Management of Capacity Building Center (CBC) contracts​

Coordinating Center​

  • Meeting scheduling/communication management​

Investigator Initiated Research​

  • 8 U54s: Serological Sciences Centers of Excellence​

  • 13 U01s: Serological Sciences Research Projects​

4 Capacity Building Centers​

  • Mt. Sinai​

  • Arizona State University

  • University of Minnesota​

  • Northwell Feinstein

FNL Developments within SeroNet​

FNL SARS-CoV-2 Serology Laboratory​

SARS-CoV-2 US Serology Standard2

  • Pooled plasma from 4 donors​

  • Contains IgG and IgM antibodies against SARS-CoV-2 Spike and Nucleocapsid​

  • Calibrated against to the World Health Organization (WHO) International Standard​

  • Validated in 8 different laboratories via:​

  • Automated Chemiluminescence​

  • Commercial Ligand Binding​

  • In-House Developed Ligand Binding​

  • Fluorescence Reduction Neutralization​

  • Distributed to >170 laboratories in US and abroad​

WHO Standards Manual3

  • Collaborated with the WHO to publish the “WHO Manual for the preparation of reference materials for use as secondary standards in antibody testing, TRS 1043, Annex 2”.​

Graph of sensitivity and specificity of COVID-19 antibody tests.
Figure 1. Sensitivity and specificity measurements commercial, in-house, and neutralization serological assays. Graphical representation of the sensitivity and specificity of all tested assays. Specificity above 93% and sensitivity above 90% were considered acceptable.

Serology Assay Comparison Study​

  • Evaluated 27 serology assays from 17 participating institutions​

  • Results normalized against WHO International Standard ​

  • Most commercial assays had high specificity (≥90%) and sensitivity (≥93%), ​

  • In-house sensitivity was ≥93% ​

  • Neutralization results were variable​

  • Manuscript submitted for publication5

Validated In-House Serology Assays​

  • ELISA for Spike and Nucleocapsid for different isotypes (IgM, IgG)​

  • Luminex-based Multiplex Assay for 18 Coronaviruses Antigens for IgG including variants and common coronaviruses (manuscript accepted for publication4)​

  • Antibody Avidity Assay for Spike IgG ​

​ImmPort

  • Worked with SeroNet PIs to obtain raw scientific data, enabling curation of 98 SeroNet publications for public access in ImmPort
  • SeroNet studies in ImmPort currently use 6 (of 15) defined research focuses and features 24 defined assay methods.
  • SeroNet studies are searchable by research focus, condition/disease, or assay method, and data files are downloadable for data mining. 
Charts of publication statistics
Figure 2. SeroNet publication demographics on ImmPort. The 98 SeroNet studies uploaded to ImmPort fall under the Cell Biology, Molecular Biology, Infection Response, Vaccine Response, and Immune Response Research Focus categories (A), while searchable assay methods include RNA sequencing, bead array assays, flow cytometry, ELISPOT, ELISA, and PCR (B).

SeroNet Developments

Capacity Building Centers

Longitudinal Studies

  • Centralized collection of annotated, well characterized serum, plasma, PBMCs and clinical data

Unique cancer cohorts

  • Numerous cancer cohorts, predominantly Multiple Myeloma
Two bar charts
Figure 3. Overview of Longitudinal Specimen Collections by Cohort graphed against days from primary vaccination

​Conclusions

  • Longitudinal serum collections from specialized populations will enable us to profile the Anti-Spike IgG over time.
  • Longitudinal PBMC collections from specialized populations will enable us to profile the Cellular Immune response over time.
  • The VICD played a key role in enabling validation of assays for SARS-CoV-2 based upon proven HPV experience.
  • Continued development of variant specific validation panels and serum standards will help the research community.
  • Infrastructure developed for SeroNet at VICD is transferrable to other diseases and future pandemic threats.
Flow chart
Figure 3.  Operational Infrastructure was developed to procure samples, test them, aliquot them and provide them for use, in Validation Panel development, to the VICD Serology Lab as well as to SeroNet investigators.

Future directions

Network capable of delivering standards and validation panels for the purpose of assay validation, enabling submission to the FDA as well as support for serosurveillance studies.
 

Reference Material Study

 

  • Develop SARS-CoV-2 Standards (plasma) and Validation Panels (serum) informed by vaccination/infection history 
  • Evaluate assays developed to detect SARS-CoV-2 for both research and clinical diagnostic purposes

Longitudinal Study

  • Collect blood samples from healthy patients and cancer patients at multiple timepoints over the course of a year
  • Characterization of the immune response to infection and vaccination

References

1. Pinto LA, Shawar RM, O'Leary B, Kemp TJ, Cherry J, Thornburg N, et al. A Trans-Governmental Collaboration to Independently Evaluate SARS-CoV-2 Serology Assays. Microbiol Spectr. 2022;10(1):e0156421.​

2. Kemp TJ, Quesinberry JT, Cherry J, Lowy DR, Pinto LA. Selection, Characterization, Calibration, and Distribution of the U.S. Serology Standard for Anti-SARS-CoV-2 Antibody Detection. J Clin Microbiol. 2022;60(11):e0099522.​

3. WHO. WHO manual for the establishment of national and other secondary standards for antibodies against infectious agents focusing on SARS-CoV2. 01/30/2022.​

4. Daisy R Roy, Troy J Kemp, Katarzyna Haynesworth, Sarah A Loftus, Ligia A Pinto. Development, Validation, and Utilization of a Luminex-Based SARS-CoV-2 Multiplex Serology Assay. Accepted for publication in ASM: Spectrum. ​

5. Troy J. Kemp, Heidi A. Hempel, Yuanji Pan, Daisy Roy, Jim Cherry, Douglas R. Lowy, Ligia A. Pinto. Assay Harmonization Study to Measure Immune Response to SARS-CoV-2 Infection and Vaccines: A Serology Methods Study.​

6. FrederickNationalLaboratory. Serology Standard Frederick National Laboratory2023 [Available from: https://frederick.cancer.gov/initiatives/seronet/serology-standard.​

Tags:  Virology