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Handbook of ELISPOT

Methods and Protocols

  • Book
  • © 2024
  • Latest edition

Overview

Editors:
  1. Alexander E. Kalyuzhny

    1. Biotechne, Minneapolis, USA

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 2768)

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Table of contents (18 protocols)

  1. Front Matter

    Pages i-xi
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  2. Important Considerations for ELISpot Validation

    • Sylvia Janetzki
    Pages 1-13

  3. Current Trends in Validating Antibody Specificities for ELISpot by Western Blotting

    • Biji T. Kurien, R. Hal Scofield
    Pages 15-27

  4. An Overview of Peptides and Peptide Pools for Antigen-Specific Stimulation in T-Cell Assays

    • Karsten Schnatbaum, Pavlo Holenya, Sebastian Pfeil, Michael Drosch, Maren Eckey, Ulf Reimer et al.
    Pages 29-50

  5. The Applications of ELISpot in the Identification and Treatment of Various Forms of Tuberculosis and in the Cancer Immunotherapies

    • Hemant K. Mishra
    Pages 51-58

  6. Artificial Intelligence-Based Counting Algorithm Enables Accurate and Detailed Analysis of the Broad Spectrum of Spot Morphologies Observed in Antigen-Specific B-Cell ELISPOT and FluoroSpot Assays

    • Alexey Y. Karulin, Melinda Katona, Zoltán Megyesi, Greg A. Kirchenbaum, Paul V. Lehmann
    Pages 59-85Open Access

  7. Comparing Flow Cytometry and ELISpot for Detection of IL-10, IL-6, and TNF Alpha on Human PBMCs

    • Kristina Boss, Jodi Hagen, Megan Constans, Christine Goetz, Alexander E. Kalyuzhny
    Pages 87-103

  8. Reagent Tracker™ Platform Verifies and Provides Audit Trails for the Error-Free Implementation of T-Cell ImmunoSpot® Assays

    • Alexander A. Lehmann, Diana R. Roen, Zoltán Megyesi, Paul V. Lehmann
    Pages 105-115Open Access

  9. Detection of SARS-CoV-2-Specific Cells Utilizing Whole Proteins and/or Peptides in Human PBMCs Using IFN-Æ´ ELISPOT Assay

    • Madeleine M. Rasche, Ella C. Kaufmann, Tamar Ratishvili, Ilya M. Swanson, Inna G. Ovsyannikova, Richard B. Kennedy
    Pages 117-133

  10. Interferon-γ/IL-2 ELISpot and mRNA Responses to the SARS-CoV2, Feline Coronavirus Serotypes 1 (FCoV1), and FCoV2 Receptor Binding Domains by the T Cells from COVID-19-Vaccinated Humans and FCoV1-Infected Cats

    • Sabarinath Nair, Bikash Sahay, Ananta P. Arukha, Lekshmi K. Edison, Chiquitha D. Crews, John G. Morris Jr et al.
    Pages 135-151

  11. Frequencies of SARS-CoV-2 Spike Protein-Specific Memory B Cells in Human PBMCs, Quantified by ELISPOT Assay

    • Ilya M. Swanson, Iana H. Haralambieva, Madeleine M. Rasche, Inna G. Ovsyannikova, Richard B. Kennedy
    Pages 153-166

  12. Monitoring Memory B Cells by Next-Generation ImmunoSpot® Provides Insights into Humoral Immunity that Measurements of Circulating Antibodies Do Not Reveal

    • Paul V. Lehmann, Zhigang Liu, Noémi Becza, Alexis V. Valente, Junbo Wang, Greg A. Kirchenbaum
    Pages 167-200Open Access

  13. Tracking Circulating HLA-Specific IgG-Producing Memory B Cells with the B-Cell ImmunoSpot Assay

    • Delphine Kervella, Sebastiaan Heidt, Robert Fairchild, Stephen Todryk, Oriol Bestard
    Pages 201-209

  14. Assessing the Affinity Spectrum of the Antigen-Specific B Cell Repertoire via ImmunoSpot®

    • Noémi Becza, Zhigang Liu, Jack Chepke, Xing-Huang Gao, Paul V. Lehmann, Greg A. Kirchenbaum
    Pages 211-239Open Access

  15. Using PBMCs in a Multiplex FluoroSpot Assay for Detection of Innate Immune Response-Modulating Impurities (IIRMIs)

    • Bartek Makower, Niklas Ahlborg
    Pages 241-249

  16. Four-Color ImmunoSpot® Assays Requiring Only 1–3 mL of Blood Permit Precise Frequency Measurements of Antigen-Specific B Cells-Secreting Immunoglobulins of All Four Classes and Subclasses

    • Lingling Yao, Noémi Becza, Andrea Maul-Pavicic, Jack Chepke, Greg A. Kirchenbaum, Paul V. Lehmann
    Pages 251-272Open Access

  17. High-Plex ELISPOT: FOLISPOT Based on Fluorescence Detection and DNA Complementary Pairing

    • Mi Liu
    Pages 273-296

  18. Triple-Color FluoroSpot Analysis of Polyfunctional Antigen-Specific T Cells by Quantification of Spot-Forming Units and Relative Spot Volumes

    • Niklas Ahlborg, Christian Smedman, Bartek Makower
    Pages 297-304

  19. Performance and Stability of New Class of Fetal Bovine Sera (FBS) and Its Lyophilized Form in ELISpot and FluoroSpot Assays: Applications for Monitoring the Immune Response in Vaccine, and Cell and Gene Immunotherapy in Clinical Trials

    • Zhinous Hosseini, Christopher J. Groves, Penny Anders, Kristen Cave, Madelyn Krunkosky, Brandi Chappell et al.
    Pages 305-316

  20. Back Matter

    Pages 317-320
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Keywords

About this book

This fourth edition volume expands on the previous editions with discussions on the latest protocols that can be applied in different laboratory landscapes ranging from purely research to clinical diagnostic environments. This volume particularly focuses on ELISpot validation and evaluation of antibodies used in this assay. The chapters in this volume cover topics such as peptides in T-Cell assays and ELISpot applications for monitoring infectious diseases like tuberculosis and SARS-CoV2; FluoroSpot; the use of AI for studying ELISpot; and comparing ELISpot with flow cytometry, feline coronavirus (FCoV2), and memory b-cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Handbook of ELISPOT: Methods and Protocols, Fourth Edition is a valuable resource to aid in the advancement of science and technology, and inspire scientists and researchers to determine and pursue new discoveries in this developing field.

Chapters 5, 7, 11, 13, and 15 are available open access under  Creative Commons Attribution 4.0 International License via link.springer.com. 

Editors and Affiliations

  • Biotechne, Minneapolis, USA

    Alexander E. Kalyuzhny

Bibliographic Information

  • Book Title: Handbook of ELISPOT

  • Book Subtitle: Methods and Protocols

  • Editors: Alexander E. Kalyuzhny

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-0716-3690-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature 2024

  • Hardcover ISBN: 978-1-0716-3689-3Published: 20 March 2024

  • Softcover ISBN: 978-1-0716-3692-3Due: 20 April 2024

  • eBook ISBN: 978-1-0716-3690-9Published: 19 March 2024

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 4

  • Number of Pages: XI, 320

  • Number of Illustrations: 1 b/w illustrations

  • Topics: Immunology

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