Development of an ultra-sensitive human IL-33 biomarker assay for age-related macular degeneration and asthma drug development

Journal of Translational Medicine

Gerald Nakamura¹  Jack Bevers¹  Kellen Schneider¹  Dhaya Seshasayee¹  Jia Wu¹  Hongkang Xi²  Laetitia Comps-Agrar³  Kelly M. Loyet³  Joyce Chan³  Elaine Mai³  Levina Goon⁴  Braeden K. Ego⁵  Menno van Lookeren Campagne⁶  Michele Grimbaldeston⁷  Vahan B. Indjeian⁷  Manda Wong⁸  Tiffany Wong⁸  Racquel Corpuz⁸ 

[1] Department of Antibody Engineering, Genentech Inc., South San Francisco, CA, USA;Department of Antibody Engineering, Genentech Inc., South San Francisco, CA, USA;Department of Immunology, Genentech Inc., South San Francisco, CA, USA;Department of Biochemical and Cellular Pharmacology, Genentech Inc., South San Francisco, CA, USA;Department of Biochemical and Cellular Pharmacology, Genentech Inc., South San Francisco, CA, USA;Department of Biology and Compound Repository, Exelixis, Alameda, CA, USA;Department of Biochemical and Cellular Pharmacology, Genentech Inc., South San Francisco, CA, USA;Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA;Department of Immunology, Genentech Inc., South San Francisco, CA, USA;Department of Inflammation and Oncology, Amgen Research, Amgen Inc., South San Francisco, CA, USA;Department of OMNI-Biomarker Development, Genentech Inc., South San Francisco, CA, USA;Department of Structural Biology, Genentech Inc., South San Francisco, CA, USA;

关键词: IL-33;    ST2;    iPCR;    Biomarker;    Age-related macular degeneration;    Asthma;   

DOI  :  10.1186/s12967-021-03189-3

来源: Springer

【 摘 要 】

BackgroundOver the past decade, human Interleukin 33 (hIL-33) has emerged as a key contributor to the pathogenesis of numerous inflammatory diseases. Despite the existence of several commercial hIL-33 assays spanning multiple platform technologies, their ability to provide accurate hIL-33 concentration measurements and to differentiate between active (reduced) and inactive (oxidized) hIL-33 in various matrices remains uncertain. This is especially true for lower sample volumes, matrices with low hIL-33 concentrations, and matrices with elevated levels of soluble Interleukin 1 Receptor-Like 1 (sST2), an inactive form of ST2 that competes with membrane bound ST2 for hIL-33 binding.ResultsWe tested the performance of several commercially available hIL-33 detection assays in various human matrices and found that most of these assays lacked the sensitivity to accurately detect reduced hIL-33 at biologically relevant levels (sub-to-low pg/mL), especially in the presence of human sST2 (hsST2), and/or lacked sufficient target specificity. To address this, we developed and validated a sensitive and specific enzyme-linked immunosorbent assay (ELISA) capable of detecting reduced and total hIL-33 levels even in the presence of high concentrations of sST2. By incorporating the immuno-polymerase chain reaction (iPCR) platform, we further increased the sensitivity of this assay for the reduced form of hIL-33 by ~ 52-fold. Using this hIL-33 iPCR assay, we detected hIL-33 in postmortem human vitreous humor (VH) samples from donors with age-related macular degeneration (AMD) and found significantly increased hIL-33 levels when compared to control individuals. No statistically significant difference was observed in aqueous humor (AH) from AMD donors nor in plasma and nasosorption fluid (NF) from asthma patients compared to control individuals.ConclusionsUnlike existing commercial hIL-33 assays, our hIL-33 bioassays are highly sensitive and specific and can accurately quantify hIL-33 in various human clinical matrices, including those with high levels of hsST2. Our results provide a proof of concept of the utility of these assays in clinical trials targeting the hIL-33/hST2 pathway.

【 授权许可】

CC BY