Measurement of IL-21 in human serum and plasma using ultrasensitive MSD S-PLEX® and Quanterix SiMoA methodologies

检出限 免疫学 医学 化学 人血浆 内科学 色谱法
作者
Josh Poorbaugh,Tanushree Samanta,S W Bright,Sean E. Sissons,Ching-Yun Chang,Pankaj Oberoi,Angus Macdonald,Andrea P. Martin,Karen L. Cox,Robert J. Benschop
出处
期刊:Journal of Immunological Methods [Elsevier BV]
卷期号:466: 9-16 被引量:19
标识
DOI:10.1016/j.jim.2018.12.005
摘要

IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commercial assays in published literature were not specific or sensitive enough to detect levels of IL-21 in heparin plasma or serum from healthy human individuals. This finding prompted an effort to develop more specific and sensitive methods to quantitate IL-21 in complex biological matrices using proprietary anti-IL-21 antibodies with the Quanterix SiMoA platform and the Meso Scale Discovery (MSD) S-PLEX® format. Assays developed on both technology platforms were characterized in heparin plasma and serum using spike recoveries across a range of concentrations. Each method was able to detect sub-pg/mL levels of IL-21 (predicted Limit of Detection [LOD] of approximately 1.0 fg/mL for both the Quanterix SiMoA and MSD S-PLEX® platforms) which is 200–500 times lower than current commercial assays. Additionally we demonstrated that rheumatoid factor did not interfere with measuring IL-21 in the Quanterix SiMoA assay. Results obtained with the two new ultrasensitive assays showed a strong correlation (r = 0.9428; p < .0001). Additionally, IL-21 levels were significantly increased in samples from patients with Systemic Lupus Erythematosus (mean+/− SD: n = 14, 202.64 +/− 111.47 fg/mL, p = .0001 for Quanterix SiMoA and 275.4 +/− 174.66 fg/mL p = .0001 for MSD S-PLEX®) as well as in samples from patients with Sjögren's Syndrome (mean+/− SD: n = 11, 122.18 +/− 84.50 fg/mL, p = .0029 for Quanterix SiMoA and 183.64 +/− 153.00 fg/mL, p = .0082 for MSD S-PLEX®) when compared to healthy donors (mean+/− SD: n = 11, 38.1 +/− 27.8 fg/mL for Quanterix SiMoA and 58.1 +/− 30.7 fg/mL for MSD S-PLEX®). These ultrasensitive assays, for the first time, allow for the accurate quantitation of human IL-21 in heparin plasma and serum. In addition, these experiments also provide a direct comparison of the MSD S-PLEX® format and Quanterix SiMoA platform technologies, which may have broader implications to future application of these methods to evaluate low abundance proteins in complex biological matrices.
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