title: An 8 minute colorimetric paper-based reverse phase vertical flow serum microarray for screening of hyper IgE syndrome.
abstract: Reverse phase microarrays are useful tools for affinity-based detection in hundreds of samples simultaneously. However, current methods typically require long assay times and fluorescent detection. Here we describe a paper-based Vertical Flow Microarray (VFM) assay as a rapid 8-minute colorimetric alternative for reverse phase microarray analysis. The VFM platform was optimized for detection of IgE with a detection limit of 1.9 μg mL(-1) in whole serum. Optimized conditions were then used to screen 113 serum samples simultaneously for hyper IgE syndrome (hIgE), a rare primary immunodeficiency characterized by elevated levels of IgE. The same set of samples were then analysed with a conventional planar microarray with fluorescent detection for head-to-head testing. Both assays found elevated levels in three out of four hIgE patient samples, whereas no control samples displayed elevated levels in either method. The comparison experiments showed a good correlation between the two assays, as determined from a linear correlation study (Pearson's r = 0.76). Further, the assay-time reduction and reproducibility (intra assay CV = 12.4 ± 4.11%) demonstrate the applicability of the VFM platform for high throughput reverse phase screening.
authors.affiliation: Division of Proteomics and Nanobiotechnology, Science for Life Laboratory, KTH Royal Institute of Technology, Sweden. helene.andersson.svahn@scilifelab.se.
authors.forename: Philippa
authors.initials: P
authors.lastname: Reuterswärd
authors.forename: Jesper
authors.initials: J
authors.lastname: Gantelius
authors.forename: Helene
authors.initials: H
authors.lastname: Andersson Svahn
mesh_terms: D019540:Area Under Curve; D003101:Collodion / Q000737:chemistry; D003124:Colorimetry / Q000295:instrumentation* / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis*; D007589:Job Syndrome / Q000097:blood* / Q000175:diagnosis*; D057230:Limit of Detection; D046228:Microarray Analysis; D010209:Paper; D040081:Protein Array Analysis / Q000379:methods; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D007073:Immunoglobulin E; D003101:Collodion