Skip to main content

Advertisement

SpringerLink
Log in

The Mystery of Antibodies Against Polyethylene Glycol (PEG) - What do we Know?

  • Research Paper
  • Published:
Pharmaceutical Research Aims and scope Submit manuscript

Abstract

Purpose

Recent findings demonstrated anti-PEG antibody formation in some healthy individuals and patients who have not received PEGylated biotherapeutics. Some of these findings evoked criticism because of shortcomings in the antibody assays used. To better understand this topic, we established robust antibody analytics and screened two cohorts of healthy individuals and one cohort of hemophilia patients for the expression of anti-PEG antibodies.

Methods

A flow cytometry approach and a fully validated ELISA platform were established to detect specific anti-PEG antibodies. Immunohistochemistry was used to test for potential binding of anti-PEG antibodies to human tissues.

Results

IgM and/or IgG anti-PEG antibodies are expressed by some healthy individuals and by some patients with hemophilia who have not received PEGylated biotherapeutics. These antibodies can be either transient or persistent and recognize PEGs of different sizes with or without terminal methoxy groups. Age and location of healthy individuals influence the prevalence of IgG but not of IgM antibodies. Anti-PEG antibodies do not cross-react with human tissues supporting the safety of the antibodies.

Conclusion

We confirm that some healthy individuals and some patients with hemophilia express specific antibodies against PEG which are not associated with any pathology and do not bind to human tissues.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

CI:

Confidence intervals

CRO:

Contract research organisation

ELISA:

Enzyme linked immunosorbent assay

FCM:

Flow cytometry

FDA:

Food and drug administration

GLP:

Good laboratory practice

IHC:

Immunohistochemistry

KA :

Equilibrium association constant

PEG:

Polyethylene glycol

PK:

Pharmacokinetic

References

  1. Polson A. A theory for the displacement of proteins and viruses with polyethylene glycol. Prep Biochem. 1977;7(2):129–54.

    CAS  PubMed  Google Scholar 

  2. Gombotz WR, Wang GH, Horbett TA, Hoffman AS. Protein adsorption to poly(ethylene oxide) surfaces. J Biomed Mater Res. 1991;25(12):1547–62.

    Article  CAS  PubMed  Google Scholar 

  3. Roberts MJ, Bentley MD, Harris JM. Chemistry for peptide and protein PEGylation. Adv Drug Deliv Rev. 2002;54(4):459–76.

    Article  CAS  PubMed  Google Scholar 

  4. Pasut G, Veronese FM. State of the art in PEGylation: the great versatility achieved after forty years of research. J Control Release. 2012;161(2):461–72.

    Article  CAS  PubMed  Google Scholar 

  5. Levy Y, Hershfield MS, Fernandez-Mejia C, Polmar SH, Scudiery D, Berger M, et al. Adenosine deaminase deficiency with late onset of recurrent infections: response to treatment with polyethylene glycol-modified adenosine deaminase. J Pediatr. 1988;113(2):312–7.

    Article  CAS  PubMed  Google Scholar 

  6. Graham ML. Pegaspargase: a review of clinical studies. Adv Drug Deliv Rev. 2003;55(10):1293–302.

    Article  CAS  PubMed  Google Scholar 

  7. Wang YS, Youngster S, Grace M, Bausch J, Bordens R, Wyss DF. Structural and biological characterization of pegylated recombinant interferon alpha-2b and its therapeutic implications. Adv Drug Deliv Rev. 2002;54(4):547–70.

    Article  CAS  PubMed  Google Scholar 

  8. Rajender Reddy K, Modi MW, Pedder S. Use of peginterferon alfa-2a (40 KD) (Pegasys) for the treatment of hepatitis C. Adv Drug Deliv Rev. 2002;54(4):571–86.

    Article  CAS  PubMed  Google Scholar 

  9. Trainer PJ, Drake WM, Katznelson L, Freda PU, Herman-Bonert V, van der Lely AJ, et al. Treatment of acromegaly with the growth hormone-receptor antagonist pegvisomant. N Engl J Med. 2000;342(16):1171–7.

    Article  CAS  PubMed  Google Scholar 

  10. Ng EW, Shima DT, Calias P, Cunningham Jr ET, Guyer DR, Adamis AP. Pegaptanib, a targeted anti-VEGF aptamer for ocular vascular disease. Nat Rev Drug Discov. 2006;5(2):123–32.

    Article  CAS  PubMed  Google Scholar 

  11. Kinstler O, Molineux G, Treuheit M, Ladd D, Gegg C. Mono-N-terminal poly(ethylene glycol)-protein conjugates. Adv Drug Deliv Rev. 2002;54(4):477–85.

    Article  CAS  PubMed  Google Scholar 

  12. Macdougall IC. CERA (Continuous Erythropoietin Receptor Activator): a new erythropoiesis-stimulating agent for the treatment of anemia. Curr Hematol Rep. 2005;4(6):436–40.

    CAS  PubMed  Google Scholar 

  13. Choy EH, Hazleman B, Smith M, Moss K, Lisi L, Scott DG, et al. Efficacy of a novel PEGylated humanized anti-TNF fragment (CDP870) in patients with rheumatoid arthritis: a phase II double-blinded, randomized, dose-escalating trial. Rheumatology (Oxford). 2002;41(10):1133–7.

    Article  CAS  Google Scholar 

  14. Sherman MR, Saifer MG, Perez-Ruiz F. PEG-uricase in the management of treatment-resistant gout and hyperuricemia. Adv Drug Deliv Rev. 2008;60(1):59–68.

    Article  CAS  PubMed  Google Scholar 

  15. Hu X, Cui Y, White J, Zhu Y, Deykin A, Nestorov I, et al. Pharmacokinetics and pharmacodynamics of peginterferon beta-1a in patients with relapsing-remitting multiple sclerosis in the randomized ADVANCE study. Br J Clin Pharmacol. 2015;79(3):514–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Konkle BA, Stasyshyn O, Chowdary P, Bevan DH, Mant T, Shima M, et al. Pegylated, full-length, recombinant factor VIII for prophylactic and on-demand treatment of severe hemophilia A. Blood. 2015;126(9):1078–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ivens IA, Baumann A, McDonald TA, Humphries TJ, Michaels LA, Mathew P. PEGylated therapeutic proteins for haemophilia treatment: a review for haemophilia caregivers. Haemophilia. 2013;19(1):11–20.

    Article  CAS  PubMed  Google Scholar 

  18. Powell JS. Lasting power of new clotting proteins. Hematol Am Soc Hematol Educ Program. 2014;2014(1):355–63.

    Google Scholar 

  19. Mannucci PM. Half-life extension technologies for haemostatic agents. Thromb Haemost. 2015;113(1):165–76.

    Article  PubMed  Google Scholar 

  20. Dreborg S, Akerblom EB. Immunotherapy with monomethoxypolyethylene glycol modified allergens. Crit Rev Ther Drug Carrier Syst. 1990;6(4):315–65.

    CAS  PubMed  Google Scholar 

  21. Ivens IA, Achanzar W, Baumann A, et al. PEGylated biopharmaceuticals: current experience and considerations for nonclinical development. Toxicol Pathol. 2015;43(7):959–83.

    Article  CAS  PubMed  Google Scholar 

  22. Armstrong JK, Hempel G, Koling S, Chan LS, Fisher T, Meiselman HJ, et al. Antibody against poly(ethylene glycol) adversely affects PEG-asparaginase therapy in acute lymphoblastic leukemia patients. Cancer. 2007;110(1):103–11.

    Article  PubMed  Google Scholar 

  23. Ganson NJ, Kelly SJ, Scarlett E, Sundy JS, Hershfield MS. Control of hyperuricemia in subjects with refractory gout, and induction of antibody against poly(ethylene glycol) (PEG), in a phase I trial of subcutaneous PEGylated urate oxidase. Arthritis Res Ther. 2006;8(1):R12.

    Article  PubMed  Google Scholar 

  24. Hershfield MS, Ganson NJ, Kelly SJ, Scarlett EL, Jaggers DA, Sundy JS. Induced and pre-existing anti-polyethylene glycol antibody in a trial of every 3-week dosing of pegloticase for refractory gout, including in organ transplant recipients. Arthritis Res Ther. 2014;16(2):R63.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Verhoef JJ, Carpenter JF, Anchordoquy TJ, Schellekens H. Potential induction of anti-PEG antibodies and complement activation toward PEGylated therapeutics. Drug Discov Today. 2014;19(12):1945–52.

    Article  CAS  PubMed  Google Scholar 

  26. Rock KL, Benacerraf B, Abbas AK. Antigen presentation by hapten-specific B lymphocytes. I. Role of surface immunoglobulin receptors. J Exp Med. 1984;160(4):1102–13.

    Article  CAS  PubMed  Google Scholar 

  27. Judge A, McClintock K, Phelps J, Maclachlan I. Hypersensitivity and loss of disease site targeting caused by antibody responses to PEGylated liposomes. Mol Ther. 2006;13(2):328–37.

    Article  CAS  PubMed  Google Scholar 

  28. Myler H, Hruska MW, Srinivasan S, Cooney E, Kong G, Dodge R, et al. Anti-PEG antibody bioanalysis: a clinical case study with PEG-IFN-λ-1a and PEG-IFN-α2a in naive patients. Bioanalysis. 2015;7(9):1093–106.

    Article  CAS  PubMed  Google Scholar 

  29. Richter AW, Akerblom E. Polyethylene glycol reactive antibodies in man: titer distribution in allergic patients treated with monomethoxy polyethylene glycol modified allergens or placebo, and in healthy blood donors. Int Arch Allergy Appl Immunol. 1984;74(1):36–9.

    Article  CAS  PubMed  Google Scholar 

  30. Armstrong JK. The occurrence, induction, specificity and potential effect of antibodies against poly(ethylene glycol). In: Veronese FM, ed. PEGylated protein drugs: basic science and clinical applications. Basel; 2009:147–168.

  31. Liu Y, Reidler H, Pan J, Milunic D, Qin D, Chen D, et al. A double antigen bridging immunogenicity ELISA for the detection of antibodies to polyethylene glycol polymers. J Pharmacol Toxicol Methods. 2011;64(3):238–45.

    Article  CAS  PubMed  Google Scholar 

  32. Fruijtier-Polloth C. Safety assessment on polyethylene glycols (PEGs) and their derivatives as used in cosmetic products. Toxicology. 2005;214(1–2):1–38.

    Article  PubMed  Google Scholar 

  33. Arora R, Srinivasan R. Is polyethylene glycol safe and effective for chronic constipation in children? Arch Dis Child. 2005;90(6):643–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zhao L, Seth A, Wibowo N, Zhao CX, Mitter N, Yu C, et al. Nanoparticle vaccines. Vaccine. 2014;32(3):327–37.

    Article  PubMed  Google Scholar 

  35. Schellekens H, Hennink WE, Brinks V. The immunogenicity of polyethylene glycol: facts and fiction. Pharm Res. 2013;30(7):1729–34.

    Article  CAS  PubMed  Google Scholar 

  36. Stidl R, Fuchs S, Bossard M, Siekmann J, Turecek PL, Putz M. Safety of PEGylated recombinant human full-length coagulation factor VIII (BAX 855) in the overall context of PEG and PEG conjugates. Haemophilia. 2016;22(1):54–64.

    Article  CAS  PubMed  Google Scholar 

  37. Wilson EB. Probable inference, the law of succession, and statistical inference. J Am Stat Assoc. 1927;158(22):209–12.

    Article  Google Scholar 

  38. Bonate PL, Sung C, Welch K, Richards S. Conditional modeling of antibody titers using a zero-inflated poisson random effects model: application to Fabrazyme. J Pharmacokinet Pharmacodyn. 2009;36(5):443–59.

    Article  CAS  PubMed  Google Scholar 

  39. Zeleis A, Kleiber C, Jackman S. Regression models for count data in R. J Stat Softw. 2008;27(8):1–25.

    Google Scholar 

  40. Saifer MG, Williams LD, Sobczyk MA, Michaels SJ, Sherman MR. Selectivity of binding of PEGs and PEG-like oligomers to anti-PEG antibodies induced by methoxyPEG-proteins. Mol Immunol. 2014;57(2):236–46.

    Article  CAS  PubMed  Google Scholar 

  41. Armstrong JK, Leger R, Wenby RB, Meiselman HJ, Garay RP, Fisher T. Occurrence of an antibody to poly(ethylene glycol) in normal donors. ASH Conference 2003.

  42. Hagen AJ, Oliver CN, Sitrin RD. Optimization of poly(ethylene glycol) precipitation of hepatitis A virus used to prepare VAQTA, a highly purified inactivated vaccine. Biotechnol Prog. 1996;12(3):406–12.

    Article  CAS  PubMed  Google Scholar 

  43. Voskuijl W, de Lorijn F, Verwijs W, Hogeman P, Heijmans J, Makel W, et al. PEG 3350 (Transipeg) versus lactulose in the treatment of childhood functional constipation: a double blind, randomised, controlled, multicentre trial. Gut. 2004;53(11):1590–4.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Reipert BM, Lubich C, Weiller M, Allacher P, Siekmann J, de la Rosa M, et al. Does PEGylated factor VIII induce antibodies against PEG?. Haemophilia 2012;18(Suppl.3):FP-MO-03.2-5.

  45. Ishida T, Wang X, Shimizu T, Nawata K, Kiwada H. PEGylated liposomes elicit an anti-PEG IgM response in a T cell-independent manner. J Control Release. 2007;122(3):349–55.

    Article  CAS  PubMed  Google Scholar 

  46. Mond JJ, Vos Q, Lees A, Snapper CM. T cell independent antigens. Curr Opin Immunol. 1995;7(3):349–54.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments and Disclosures

The authors are grateful to Julia Stevens (Asterand Bioscience) for conducting the human tissue cross-reactivity study. Moreover, we thank Neriman Yildiz, Margit Pichler and Silvia Neppl for technical assistance. The authors also thank Karima Benamara for editing the manuscript.

Contribution

C. L. designed research, performed experiments, analyzed and interpreted data, and wrote the manuscript; P.A. and F.M.H. established validated assay formats, and analyzed and interpreted data; M.d.l.R. designed and interpreted data from the tissue cross-reactivity study; A.B. performed statistical modeling of antibody titers; T.P. performed experiments and interpreted data; J.S. generated PEGylated proteins, designed and interpreted data from the prevalence study; J.O. provided patient samples and interpreted data; F.S. interpreted data; B.M.R. designed research, analyzed and interpreted data, and wrote the manuscript.

Conflict-of-interest disclosure: C. L., M.d.l.R., A.B., T.P., F.M.H., J.S., H.P.S., F.S. and B.M.R. are employees of Baxalta Innovations GmbH. P.A. was an employee of Baxter Innovation GmbH in the course of this study and is now employed by IMC FH Krems, University for Applied Sciences, Krems, Austria.

Author information

Authors and Affiliations

  1. Baxalta Innovations GmbH, Industriestrasse 72, A-1220, Vienna, Austria

    Christian Lubich, Maurus de la Rosa, Alexander Bauer, Thomas Prenninger, Frank Michael Horling, Jürgen Siekmann, Friedrich Scheiflinger & Birgit Maria Reipert

  2. IMC FH Krems, University for Applied Sciences, Krems, Austria

    Peter Allacher

  3. Institute for Experimental Hematology and Transfusion Medicine, Bonn, Germany

    Johannes Oldenburg

Authors
  1. Christian Lubich
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Allacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maurus de la Rosa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Alexander Bauer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Thomas Prenninger
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Frank Michael Horling
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jürgen Siekmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Johannes Oldenburg
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Friedrich Scheiflinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Birgit Maria Reipert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Birgit Maria Reipert.

Additional information

This study was supported by Baxalta Innovations GmbH.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(DOCX 45 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lubich, C., Allacher, P., de la Rosa, M. et al. The Mystery of Antibodies Against Polyethylene Glycol (PEG) - What do we Know?. Pharm Res 33, 2239–2249 (2016). https://doi.org/10.1007/s11095-016-1961-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11095-016-1961-x

KEY WORDS

Search

Navigation