The AAPS Journal

, Volume 16, Issue 2, pp 194–205 | Cite as

Assay Formats: Recommendation for Best Practices and Harmonization from the Global Bioanalysis Consortium Harmonization Team

  • Sherri Dudal
  • Daniel Baltrukonis
  • Rebecca Crisino
  • M. Jaya Goyal
  • Alison Joyce
  • Karolina Österlund
  • John Smeraglia
  • Yoshitaka Taniguchi
  • Jihong Yang
Review Article Theme: Best Practices for Bioanalytical Methods: Recommendations from the Global Bioanalysis Consortium
Part of the following topical collections:
  1. Theme: Best Practices for Bioanalytical Methods: Recommendations from the Global Bioanalysis Consortium


As part of the GBC (Global Bioanalysis Consortium), the L3 assay format team has focused on reviewing common platforms used to support ligand binding assays in the detection of biotherapeutics. The following review is an overview of discussions and presentations from around the globe with a group of experts from different companies to allow an international harmonization of common practices and suggestions for different platforms. Some of the major platforms include Gyrolab, Erenna, RIA, AlphaLISA, Delfia, Immuno-PCR, Luminex, BIAcore, and ELISAs. The review is meant to support bioanalysts in taking decisions between different platforms depending on the needs of the analyte with a number of recommendations to help integration of platforms into a GLP environment.


assay format biotherapeutic large molecule platform technology 



We would like to acknowledge the input of Katherine Mckay (Covance, UK) and Mahesh Kumar (Biocon, India) in the early team sessions and their contribution to discussions.

Conflict of Interest

A possible conflict of interest would be the role of Karolina Österland in terms of Gyrolab input, but as this was a group-driven workstream and feedback from the international community, her opinion was mixed amongst the opinion of the LBA community.


This article is representative of the expert opinions of the co-authors and the international community in the field of large molecule bioanalysis. It does not represent a single company viewpoint.


  1. 1.
    Myler HA, Given A, Kolz K, Mora JR, Hristopoulos G. Biotherapeutic bioanalysis: a multi-indication case study review. Bioanalysis. 2011;3(6):623–43 [Review].PubMedCrossRefGoogle Scholar
  2. 2.
    US Department of Health and Human Services, Food and Drug administration, CDER, CVM. Guidance for Industry, Bioanalytical Method Validation, 2001.Google Scholar
  3. 3.
    DeSilva B, Smith W, Weiner R, Kelley M, Smolec J, Lee B, et al. Recommendations for the bioanalytical method validation of ligand-binding assays to support pharmacokinetic assessments of macromolecules. Pharm Res. 2003;20(11):1885–900 [Guideline].PubMedCrossRefGoogle Scholar
  4. 4.
    Fast DM, Kelley M, Viswanathan CT, O’Shaughnessy J, King SP, Chaudhary A, et al. Workshop report and follow-up–AAPS Workshop on current topics in GLP bioanalysis: assay reproducibility for incurred samples—implications of Crystal City recommendations. AAPS J. 2009;11(2):238–41 [Review].PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Smolec J, DeSilva B, Smith W, Weiner R, Kelly M, Lee B, et al. Bioanalytical method validation for macromolecules in support of pharmacokinetic studies. Pharm Res. 2005;22(9):1425–31 [Review Validation Studies].PubMedCrossRefGoogle Scholar
  6. 6.
    Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, et al. Quantitative bioanalytical methods validation and implementation: best practices for chromatographic and ligand binding assays. Pharm Res. 2007;24(10):1962–73 [Consensus Development Conference].PubMedCrossRefGoogle Scholar
  7. 7.
    European Medicines Agency. Guideline on Bioanalytical Method Validation, 2009.Google Scholar
  8. 8.
    Leary BA, Lawrence-Henderson R, Mallozzi C, Fernandez Ocana M, Duriga N, O’Hara DM, et al. Bioanalytical platform comparison using a generic human IgG PK assay format. J Immunol Methods. 2013;397(1–2):28–36.PubMedCrossRefGoogle Scholar
  9. 9.
    Soderstrom CI, Spriggs FP, Song W, Burrell S. Comparison of four distinct detection platforms using multiple ligand binding assay formats. J Immunol Methods. 2011;371(1–2):106–13 [Comparative Study Evaluation Studies].PubMedCrossRefGoogle Scholar
  10. 10.
    Forster RJ, Bertoncello P, Keyes TE. Electrogenerated chemiluminescence. Annu Rev Anal Chem (Palo Alto Calif). 2009;2:359–85 [Review].CrossRefGoogle Scholar
  11. 11.
    Lewkowich IP, Campbell JD, HayGlass KT. Comparison of chemiluminescent assays and colorimetric ELISAs for quantification of murine IL-12, human IL-4 and murine IL-4: chemiluminescent substrates provide markedly enhanced sensitivity. J Immunol Methods. 2001;247(1–2):111–8 [Comparative Study Research Support, Non-U.S. Gov’t].PubMedCrossRefGoogle Scholar
  12. 12.
    Sloan JH, Ackermann BJ, O’Dell M, Bowsher RR, Dean RA, Konrad RJ. Development of a novel radioimmunoassay to detect autoantibodies to amyloid beta peptides in the presence of a cross-reactive therapeutic antibody. J Pharm Biomed Anal. 2011;56(5):1029–34 [Research Support, Non-U.S. Gov’t Validation Studies].PubMedCrossRefGoogle Scholar
  13. 13.
    Sickert D, editor. AlphaLISA - a “no wash” high-throughput alternative to ELISA for PK, PD and immunogenicity measurement during drug development? European Bioanalytical Forum, 3rd Focus Meeting; 2012; Brussels, Belgium.Google Scholar
  14. 14.
    Fraley KJ, Abberley L, Hottenstein CS, Ulicne JJ, Citerone DR, Szapacs ME. The Gyrolab immunoassay system: a platform for automated bioanalysis and rapid sample turnaround. Bioanalysis. 2013;5(14):1765–74 [Research Support, Non-U.S. Gov’t Validation Studies].PubMedCrossRefGoogle Scholar
  15. 15.
    Liu R, Pillutla R, DeSilva B, Zhang YJ. Rapid development of multiple ‘fit-for-purpose’ assays on an automatic microfluidic system using a streamlined process in support of early biotherapeutics discovery programs. Bioanalysis. 2013;5(14):1751–63 [Research Support, Non-U.S. Gov’t].PubMedCrossRefGoogle Scholar
  16. 16.
    Liu XF, Wang X, Weaver RJ, Calliste L, Xia C, He YJ, et al. Validation of a Gyrolab assay for quantification of rituximab in human serum. J Pharmacol Toxicol Methods. 2012;65(3):107–14.PubMedCrossRefGoogle Scholar
  17. 17.
    Mora JR, Obenauer-Kutner L, Vimal PV. Application of the Gyrolab platform to ligand-binding assays: a user’s perspective. Bioanalysis. 2010;2(10):1711–5.PubMedCrossRefGoogle Scholar
  18. 18.
    Patel V, Leach D, Hornberger M, Williams K, Shih J, Ma M, et al. Automating bioanalytical sample analysis through enhanced system integration. Bioanalysis. 2013;5(13):1649–59 [Research Support, Non-U.S. Gov’t].PubMedCrossRefGoogle Scholar
  19. 19.
    St Ledger K, Agee SJ, Kasaian MT, Forlow SB, Durn BL, Minyard J, et al. Analytical validation of a highly sensitive microparticle-based immunoassay for the quantitation of IL-13 in human serum using the Erenna immunoassay system. J Immunol Methods. 2009;350(1–2):161–70 [Comparative Study].PubMedCrossRefGoogle Scholar
  20. 20.
    Todd J, Freese B, Lu A, Held D, Morey J, Livingston R, et al. Ultrasensitive flow-based immunoassays using single-molecule counting. Clin Chem. 2007;53(11):1990–5.PubMedCrossRefGoogle Scholar
  21. 21.
    Dyleski L J, A., et al., editor. Method development and validation of an alternative immunoassay platform for PK studies in a regulated environment. AAPS NBC; 2011; San Francisco, CA, USA.Google Scholar
  22. 22.
    Tarasow TM, Penny L, Patwardhan A, Hamren S, McKenna MP, Urdea MS. Microfluidic strategies applied to biomarker discovery and validation for multivariate diagnostics. Bioanalysis. 2011;3(19):2233–51 [Research Support, Non-U.S. Gov’t Review].PubMedCrossRefGoogle Scholar
  23. 23.
    Huber R, Eisenbraun J, Miletzki B, Adler M, Scheer R, Klein R, et al. Pharmacokinetics of natural mistletoe lectins after subcutaneous injection. Eur J Clin Pharmacol. 2010;66(9):889–97 [Clinical Trial, Phase I Research Support, Non-U.S. Gov’t].PubMedCrossRefGoogle Scholar
  24. 24.
    Spriggs F, Aggarwal P, Deng S, Soderstrom C, Donovan C. Comparison of an antibody capture and a cell capture ligand-binding assay to quantify a monoclonal therapeutic in serum. Bioanalysis. 2011;3(6):605–11 [Comparative Study Evaluation Studies].PubMedCrossRefGoogle Scholar
  25. 25.
    Dula M. ea, editor. Development and validation of a pharmacokinetic assay on the Gyrolab platform for use in phase II/III clinical studies. AAPS NBC; 2012; San Diego, CA, USA.Google Scholar
  26. 26.
    Lee JW, Kelley M, King LE, Yang J, Salimi-Moosavi H, Tang MT, et al. Bioanalytical approaches to quantify “total” and “free” therapeutic antibodies and their targets: technical challenges and PK/PD applications over the course of drug development. Aaps J. 2011;13(1):99–110 [Review].PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Lofgren JA, Dhandapani S, Pennucci JJ, Abbott CM, Mytych DT, Kaliyaperumal A, et al. Comparing ELISA and surface plasmon resonance for assessing clinical immunogenicity of panitumumab. J Immunol. 2007;178(11):7467–72 [Comparative Study Controlled Clinical Trial Validation Studies].PubMedCrossRefGoogle Scholar

Copyright information

© American Association of Pharmaceutical Scientists 2013

Authors and Affiliations

  • Sherri Dudal
    • 1
    • 2
  • Daniel Baltrukonis
    • 3
  • Rebecca Crisino
    • 4
  • M. Jaya Goyal
    • 5
  • Alison Joyce
    • 6
  • Karolina Österlund
    • 7
  • John Smeraglia
    • 8
  • Yoshitaka Taniguchi
    • 9
  • Jihong Yang
    • 10
  1. 1.Novartis Pharma AGNovartis Institute for Biomedical ResearchBaselSwitzerland
  2. 2.F. Hoffmann-La Roche LtdBaselSwitzerland
  3. 3.Pfizer Inc.GrotonUSA
  4. 4.Janssen, Research & Development, LLCRadnorUSA
  5. 5.Biogen IdecCambridgeUSA
  6. 6.Pfizer Inc.AndoverUSA
  7. 7.Gyros ABUppsalaSweden
  8. 8.UCB Parma S.ABraine-l’AlleudBelgium
  9. 9.Toray Research Center, Inc.Kamakura-shiJapan
  10. 10.GenentechSouth San FranciscoUSA

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