A scalable platform for highly-multiplexed analysis of antibody reactivity from <1uL of blood

Information

  • Research Project
  • 10265659
  • ApplicationId
    10265659
  • Core Project Number
    U24AI152172
  • Full Project Number
    3U24AI152172-02S1
  • Serial Number
    152172
  • FOA Number
    PA-18-591
  • Sub Project Id
  • Project Start Date
    6/29/2020 - 3 years ago
  • Project End Date
    3/31/2023 - a year ago
  • Program Officer Name
    PEYMAN, JOHN A
  • Budget Start Date
    4/1/2021 - 3 years ago
  • Budget End Date
    3/31/2022 - 2 years ago
  • Fiscal Year
    2021
  • Support Year
    02
  • Suffix
    S1
  • Award Notice Date
    4/16/2021 - 3 years ago

A scalable platform for highly-multiplexed analysis of antibody reactivity from <1uL of blood

PROJECT SUMMARY Antibodies are highly-specific, diverse and widely-assayed biomarkers used to determine recent or historical pathogen exposures, measure the protection conferred by a vaccine, understand the basis of autoimmune diseases or evaluate a host's immunological function. Traditional assays for antibodies focus on one or a small number of reactivities at a time, and so are incommensurate with the scale and diversity of an individual's antibody response. A tool to more holistically interrogate this diversity of reactivities using a small sample volume would enable a new generation of studies in systems immunology, disease association, and epidemiological surveillance. Here, we propose to optimize and significantly extend an approach we have developed for highly- multiplexed, reproducible and inexpensive assays that enable sensitive and high-resolution analysis of antibody reactivity across 100,000s of antigens from <1µL of blood. Our approach takes advantage of a rapid, fully-in- vitro method for generating 100,000s of DNA-barcoded peptides (`PepSeq') as probes for the highly-multiplexed interrogation of serum antibodies using DNA sequencing. As a proof-of-concept, we will be focusing here on an assay targeting all viruses known to infect humans (i.e., the human virome). The virome is an ideal use case for this technology, as viruses represent an incredibly diverse and ubiquitous challenge to the immune system, and because of their small genome sizes, the complete virome can be covered within a single library with minimal loss of diversity. Our preliminary data with this virome assay establishes the feasibility of this approach. Here, we will optimize the assay procedures for multiple sample types in order to increase sensitivity and specificity, while decreasing cost. We will also establish standardized protocols for isotype-specific profiling, adapt the technology to enable antigen-specific, single-cell characterization, and build a suite of open access data analysis and visualization tools to facilitate the use of this technology by the broader research community. Throughout this process, we will generate a panel of anti-virome antibody profiles, including a cohort profiled longitudinally ? this data will be made available to the community through the ImmPort portal. If successful, this project will deliver: (i) an optimized assay SOP and library for comprehensive evaluation of pan-viral immunity using a small sample volume, (ii) a large set of publicly-available anti-virome immunity datasets, and (iii) a framework for multiplexed serological assay development that can be directly extended to other targets.

IC Name
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
  • Activity
    U24
  • Administering IC
    AI
  • Application Type
    3
  • Direct Cost Amount
    223296
  • Indirect Cost Amount
    114638
  • Total Cost
    337934
  • Sub Project Total Cost
  • ARRA Funded
    False
  • CFDA Code
    855
  • Ed Inst. Type
  • Funding ICs
    NIAID:337934\
  • Funding Mechanism
    OTHER RESEARCH-RELATED
  • Study Section
    ZAI1
  • Study Section Name
    Special Emphasis Panel
  • Organization Name
    TRANSLATIONAL GENOMICS RESEARCH INST
  • Organization Department
  • Organization DUNS
    118069611
  • Organization City
    PHOENIX
  • Organization State
    AZ
  • Organization Country
    UNITED STATES
  • Organization Zip Code
    850042274
  • Organization District
    UNITED STATES