A generic method for the detection of polyethylene glycol specific IgG and IgM antibodies in human serum.

Autor: Ehlinger C; Biologics Development Sciences, Janssen BioTherapeutics, Janssen R&D, LLC, 1400 McKean Road, PO Box 776, Spring House, PA 19477, USA. Electronic address: cehlinge@its.jnj.com., Spear N; Biologics Development Sciences, Janssen BioTherapeutics, Janssen R&D, LLC, 1400 McKean Road, PO Box 776, Spring House, PA 19477, USA., Doddareddy R; Biologics Development Sciences, Janssen BioTherapeutics, Janssen R&D, LLC, 1400 McKean Road, PO Box 776, Spring House, PA 19477, USA., Shankar G; Biologics Development Sciences, Janssen BioTherapeutics, Janssen R&D, LLC, 1400 McKean Road, PO Box 776, Spring House, PA 19477, USA., Schantz A; Biologics Development Sciences, Janssen BioTherapeutics, Janssen R&D, LLC, 1400 McKean Road, PO Box 776, Spring House, PA 19477, USA.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2019 Nov; Vol. 474, pp. 112669. Date of Electronic Publication: 2019 Oct 12.
DOI: 10.1016/j.jim.2019.112669
Abstrakt: Detection of anti-drug antibodies is a critical step in the development of large molecule biopharmaceuticals. In the case of multicomponent/multifunctional molecules, such as fusion proteins and protein conjugates such as covalent polyethylene glycol (PEG)~protein conjugates, it is useful to further characterize anti-drug antibody (ADA) binding to key domains of the drug. The detection of anti-PEG antibodies poses special challenges that if overlooked can result in underreporting antibody responses. Here we describe the development and characterization of a novel ELISA to detect anti-PEG antibodies that provides a more complete interpretation of anti-PEG than other published methods. Being specific to the PEG moiety alone, this method is intended to detect anti-PEG antibodies independent of the protein to which PEG is conjugated. Based upon early indications that our assay could detect anti-PEG antibodies at a surprisingly high frequency in the general population, our emphasis throughout method development and validation was to ensure that non-specific signals and unintended interactions were not falsely contributing to detection of anti-PEG antibodies. Techniques, including orthogonal methods used to ensure that this ELISA detected antibodies specific to PEG included competition, immunodepletion, immunoprecipitation/western blot and an Octet kinetic binding analysis. The validated ELISA can detect 100 ng/mL of an anti-PEG IgG positive control and 800 ng/mL of an anti-PEG IgM positive control in the presence of 7.5 μg/mL of the PEGylated therapeutic (MW 64 kDa). The intra-assay percent co-efficient of variation (CV) and inter-assay CV of the low positive control samples in the screening method were 4.1 to 7.2% and 16.7 to 17.7%, respectively. Additional assay performance parameters that were validated are also described. When the validated assay was applied to a population of 200 healthy blood donors with no known exposure to biopharmaceutical PEG conjugates it indicated a pre-existing anti-PEG antibody prevalence of 97.5%. We suggest this surprising result is a consequence of exposure to PEG additives in everyday products, such as cosmetics, processed foods and over-the-counter (OTC) pharmaceuticals.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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