title: Development and validation of panoptic Meso scale discovery assay to quantify total systemic interleukin-6.
abstract: OBJECTIVE
  Interleukin-6 (IL-6), a multifunctional cytokine, exists in several forms ranging from a low molecular weight (MW 20-30 kDa) non-complexed form to high MW (200-450 kDa), complexes. Accurate baseline IL-6 assessment is pivotal to understand clinical responses to IL-6-targeted treatments. Existing assays measure only the low MW, non-complexed IL-6 form. The present work aimed to develop a validated assay to measure accurately total IL-6 (complexed and non-complexed) in serum or plasma as matrix in a high throughput and easily standardized format for clinical testing.
  
  
  METHODS
  Commercial capture and detection antibodies were screened against humanized IL-6 and evaluated in an enzyme-linked immunosorbent assay format. The best antibody combinations were screened to identify an antibody pair that gave minimum background and maximum recovery of IL-6 in the presence of 100% serum matrix. A plate-based total IL-6 assay was developed and transferred to the Meso Scale Discovery (MSD) platform for large scale clinical testing.
  
  
  RESULTS
  The top-performing antibody pair from 36 capture and four detection candidates was validated on the MSD platform. The lower limit of quantification in human serum samples (n = 6) was 9.77 pg l(-1) , recovery ranged from 93.13-113.27%, the overall pooled coefficients of variation were 20.12% (inter-assay) and 8.67% (intra-assay). High MW forms of IL-6, in size fractionated serum samples from myelodysplastic syndrome and rheumatoid arthritis patients, were detected by the assay but not by a commercial kit.
  
  
  CONCLUSIONS
  This novel panoptic (sees all forms) IL-6 MSD assay that measures both high and low MW forms may have clinical utility.
authors.affiliation: Oncology, Translational Research, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.
authors.forename: Shalini
authors.initials: S
authors.lastname: Chaturvedi
authors.affiliation: Oncology, Discovery, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.
authors.forename: Derick
authors.initials: D
authors.lastname: Siegel
authors.affiliation: Immunology Development, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania, USA.
authors.forename: Carrie L
authors.initials: CL
authors.lastname: Wagner
authors.affiliation: Oncology, Translational Research, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.
authors.forename: Jaehong
authors.initials: J
authors.lastname: Park
authors.affiliation: Oncology Development, Janssen Pharmaceutical Companies of Johnson & Johnson, Beerse, Belgium.
authors.forename: Helgi
authors.initials: H
authors.lastname: van de Velde
authors.affiliation: Oncology Development, Janssen Pharmaceutical Companies of Johnson & Johnson, Leiden, the Netherlands.
authors.forename: Jessica
authors.initials: J
authors.lastname: Vermeulen
authors.affiliation: Oncology, Janssen Pharmaceutical Companies of Johnson & Johnson, Raritan, New Jersey.
authors.forename: Man-Cheong
authors.initials: MC
authors.lastname: Fung
authors.affiliation: Oncology, Translational Research, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.
authors.forename: Manjula
authors.initials: M
authors.lastname: Reddy
authors.affiliation: Oncology, Translational Research, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.|Current address: MedImmune LLC, One MedImmune Way, Gaithersburg, Maryland, USA.
authors.forename: Brett
authors.initials: B
authors.lastname: Hall
authors.affiliation: Oncology, Translational Research, Janssen Pharmaceutical Companies of Johnson & Johnson, Spring House, Pennsylvania.
authors.forename: Kate
authors.initials: K
authors.lastname: Sasser
mesh_terms: D001172:Arthritis, Rheumatoid / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D015850:Interleukin-6 / Q000097:blood*; D057230:Limit of Detection; D009190:Myelodysplastic Syndromes / Q000097:blood; D012680:Sensitivity and Specificity
chemical_list: C508600:IL6 protein, human; D015850:Interleukin-6
keywords: cancer; interleukin-6; panoptic assay; rheumatoid arthritis
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