Biological drug products may elicit an antidrug antibody (ADA) response. The current widely used bridging ligand binding assay (LBA) is the gold standard for ADA assessments in drug development, which is a qualitative assay followed by a quasi-quantitative titer analysis but can be prone to interferences from biological matrices, drug targets and circulating drugs. We present our perspectives and findings in exploring a hybrid LBA/LC–MS as an orthogonal bioanalytical tool for clinical immunogenicity assessments. The hybrid LBA/LC–MS is a semiquantitative assay with acceptable specificity, drug tolerance and the capability of multiplexed detection of ADA isotypes. The assay results suggest this technology to be a promising and complementary bioanalytical tool that can provide informative immunogenicity data in drug development.

Keywords:

Papers of special note have been highlighted as: • of interest; •• of considerable interest

References

1. Mire-Sluis AR, Barrett YC, Devanarayan V et al. Recommendations for the design and optimization of immunoassays used in the detection of host antibodies against biotechnology products. J. Immunol. Methods 289, 1–16 (2004). • A commonly referenced paper for ligand binding assay (LBA) immunogenicity assay design and optimization.
Medline CASGoogle Scholar
2. Shankar G, Devanarayan V, Amaravadi L et al. Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J. Pharm. Biomed. Anal. 48(5), 1267–1281 (2008).
Medline CASGoogle Scholar
3. Shankar G, Devanarayan V, Amaravadi L et al. Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J. Pharm. Biomed. Anal. 48(5), 1267–1281 (2008).
Medline CASGoogle Scholar
4. European Medicines Agency. Guideline on immunogenicity assessment of monoclonal antibodies intended for in vivo clinical use (2012). www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2012/06/WC500128688.pdf
Google Scholar
5. Zhong ZD, Clements-Egan A, Gorovits B et al. Drug target interference in immunogenicity assays: recommendations and mitigation strategies. AAPS J. 19(6), 1564–1575 (2017).
Medline CASGoogle Scholar
6. Hart MH, Vrieze H, Wouters D et al. Differential effect of drug interference in immunogenicity assays. J. Immunol. Methods 372(1–2), 196–203 (2011).
Medline CASGoogle Scholar
7. Chen K, Page JG, Schwartz AM et al. False-positive immunogenicity responses are caused by CD20⁺ B cell membrane fragments in an anti-ofatumumab antibody bridging assay. J. Immunol. Methods 394(1–2), 22–31 (2013).
Medline CASGoogle Scholar
8. Lofgren JA, Dhandapani S, Pennucci JJ et al. Comparing ELISA and surface plasmon resonance for assessing clinical immunogenicity of panitumumab. J. Immunol. 178(11), 7467–7472 (2007).
Medline CASGoogle Scholar
9. Li J, Schantza A, Schwegler M et al. Detection of low-affinity antidrug antibodies and improved drug tolerance in immunogenicity testing by Octet^® biolayer interferometry. J. Pharm. Biomed. Anal. 54(2), 286–294 (2011).
Medline CASGoogle Scholar
10. Bourdage J, Cook CA, Farrington DL et al. An Affinity Capture Elution (ACE) assay for detection of antidrug antibody to monoclonal antibody therapeutics in the presence of high levels of drug. J. Immunol. Methods 327, 10–17 (2007).
Medline CASGoogle Scholar
11. Smith HW, Butterfield A, Sun D. Detection of antibodies against therapeutic proteins in the presence of residual therapeutic protein using a solid-phase extraction with acid dissociation (SPEAD) sample treatment prior to ELISA. Regul. Toxicol. Pharmacol. 49(3), 230–237 (2007).
Medline CASGoogle Scholar
12. Zoghbi J, Xu Y, Grabert R et al. A breakthrough novel method to resolve the drug and target interference problem in immunogenicity assays. J. Immunol. Methods. 426, 62–69 (2015).
Medline CASGoogle Scholar
13. Xu W, Jiang H, Titsch C et al. Development and characterization of a pre-treatment procedure to eliminate human monoclonal antibody therapeutic drug and matrix interference in cell-based functional neutralizing antibody assays. J. Immunol. Methods 1759(416), 318–314 (2014).
Google Scholar
14. Niu H, Klem T, Yang J et al. A biotin-drug extraction and acid dissociation (BEAD) procedure to eliminate matrix and drug interference in a protein complex antidrug antibody (ADA) isotype specific assay. J. Immunol. Methods 446, 30–36 (2017).
Medline CASGoogle Scholar
15. Carrasco-Triguero M, Mahood C, Milojic-Blair M et al. Overcoming soluble target interference in an anti-therapeutic antibody screening assay for an antibody-drug conjugate therapeutic. Bioanalysis 4(16), 2013–2026 (2012).
CASGoogle Scholar
16. Zeng J, Jiang H, Luo L. Semiquantification and isotyping of antidrug antibodies by immunocapture-LC/MS for immunogenicity assessment. Lee MSJi QC (Eds). Protein Analysis Using Mass Spectrometry: Accelerating Protein Biotherapeutics from Lab to Patient. John Wiley & Sons, Inc., NJ, USA 91–97 (2017).
Google Scholar
17. Lee JW. Generic method approaches for monoclonal antibody therapeutics analysis using both ligand binding and LC–MS/MS techniques. Bioanalysis 8(1), 19–27 (2016).
CASGoogle Scholar
18. Chappell DL, Lassman ME, McAvoy T et al. Quantitation of human peptides and proteins via MS: review of analytically validated assays. Bioanalysis 6(13), 1843–1857 (2014).
CASGoogle Scholar
19. Wilffert D, Bischoff R, van de Merbel NC. Antibody-free workflows for protein quantification by LC–MS/MS. Bioanalysis 7(6), 763–779 (2015).
CASGoogle Scholar
20. Jiang H, Zeng J, Titsch C et al. Fully validated LC–MS/MS assay for the simultaneous quantitation of co-administered therapeutic antibodies in cynomolgus monkey serum. Anal. Chem. 85(20), 9859–9867 (2013).
Medline CASGoogle Scholar
21. Furlong MT, Titsch C, Xu W et al. Evaluation of a universal LC–MS/MS assay for bioanalysis of human IgG4 subclass monoclonal antibody protein drugs and drug candidates in clinical studies. Bioanalysis 6, 1747–1758 (2014).
CASGoogle Scholar
22. Jiang H, Xu W, Titsch C et al. Innovative use of LC–MS/MS for simultaneous quantitation of neutralizing antibody, residual drug and human immunoglobulin G in immunogenicity assay development. Anal. Chem. 86(5), 2673–2680 (2014). •• A proof-of-concept report to demonstrate that hybrid LBA/LC–MS can detect antidrug antibody.
Medline CASGoogle Scholar
23. Ackermann B, Neubert H, Hughes N et al. 2015 white paper on recent issues in bioanalysis: focus on new technologies and biomarkers (Part 2 – hybrid LBA/LCMS and input from regulatory agencies). Bioanalysis 7(23), 3019–3034 (2015).
CASGoogle Scholar
24. Song A, Lee A, Garofolo F et al. 2016 white paper on recent issues in bioanalysis: focus on biomarker assay validation (BAV): (Part 2 – Hybrid LBA/LCMS and input from regulatory agencies). Bioanalysis 8(23), 2457–2474 (2016).
CASGoogle Scholar
25. Neubert H, Song A, Lee A et al. 2017 white paper on recent issues in bioanalysis: rise of hybrid LBA/LCMS immunogenicity assays (Part 2: hybrid LBA/LCMS biotherapeutics, biomarkers & immunogenicity assays and regulatory agencies’ inputs). Bioanalysis 9(23), 1895–1912 (2017).
CASGoogle Scholar
26. Chen LZ, Roos D, Philip E. Development of immunocapture-LC/MS assay for simultaneous ADA isotyping and semiquantitation. J. Immunol. Res. 2016, 7682472 (2016). •• Application of hybrid LBA/LC–MS in antidrug antibody isotyping and semiquantitation.
MedlineGoogle Scholar
27. Neubert H, Grace C, Rumpel K et al. Assessing immunogenicity in the presence of excess protein therapeutic using immunoprecipitation and quantitative mass spectrometry. Anal. Chem. 80(18), 6907–6914 (2008). • First application of indirect LBA/MS to assess immunogenicity.
Medline CASGoogle Scholar
28. Dai S, Schantz A, Clements-Egan A et al. Development of a method that eliminates false-positive results due to nerve growth factor interference in the assessment of fulranumab immunogenicity. AAPS J. 16(3), 464–477 (2014).
Medline CASGoogle Scholar
29. Nesman S, Gokemeijer J, Terryberry J et al. Isotyping therapeutic protein immunogenicity in an automated multiplexed assay. http://sqidiagnostics.com/sites/default/files/resources/pdfs/Isotyping%20Therapeutic%20Protein%20Immunogenicity%20in%20an%20Automated%20Multiplexed%20Assay.pdf
Google Scholar
30. Postelnek J, Myler H, Santockyte R et al. MSD U-PLEX^® technology differentially measures domain-specific antidrug antibodies. http://abstracts.aaps.org/Verify/NBC15/PosterSubmissions/M1030.pdf
Google Scholar
31. GlobalRPH. http://www.globalrph.com/labs_i.htm#
Google Scholar
32. Song S, Yang L, Trepicchio WL et al. Understanding the supersensitive antidrug antibody assay: unexpected high antidrug antibody incidence and its clinical relevance. J. Immunol Methods 2016,3072586 (2016).
Google Scholar
33. U.S. Department of Health and Human Services Food and Drug Administration. Immunogenicity testing of therapeutic protein products – developing and validating assays for anti-drug antibody detection (2019). www.fda.gov/media/119788/download
Google Scholar
34. Neubert H, Song A, Lee A et al. White paper: rise of hybrid LBA/LCMS immunogenicity assays (Part 2: hybrid LBA/LCMS biotherapeutics, biomarkers & immunogenicity assays and regulatory agencies’ inputs). Bioanalysis 9(23), 1895–1912 (2017). •• White paper to summarize consensus and regulatory perspectives on hybrid LBA/LC–MS immunogenicity assays.
CASGoogle Scholar

Vol. 11, No. 17

Metrics

Downloaded 940 times

History

Received 20 April 2018

Accepted 16 April 2019

Published online 18 June 2019

Published in print September 2019

Information

Keywords

Financial & competing interests disclosure

The authors have no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

PDF download

Perspectives on exploring hybrid LBA/LC–MS approach for clinical immunogenicity testing

Abstract

References