title: Feasibility of a simple microsieve-based immunoassay platform.
abstract: The intrinsic properties of silicon microsieves, such as an optically flat surface, high overall porosity, and low flow resistance have led to an increasing number of biotechnology applications. In this report, the feasibility of creating a microsieve-based immunoassay platform was explored. Microsieves containing 5μm pores were coupled with poly-acrylic acid chains, and then mounted into a plastic holder to enable rapid reagent exchanges via a wicking mechanism. The mounted microsieves were coated with infectious disease-related antigens at [2.5 and 25μg/mL], [20 and 50μg/mL], and [20 and 100μg/mL] to facilitate detection of serum-derived human antibodies against Rubella (3-day measles), B. burgdorferi (Lyme disease), or T. pallidum (syphilis), respectively. The prototype microsieve-based immunoassay platform was able to distinguish positive control sera containing antibodies against Rubella, T. pallidum, and B. burgdorferi from negative control sera with similar qualitative results as FDA-approved ELISA tests. Testing of a WHO IgG syphilitic standard at 0.3, 0.15, 0.075, 0.0375, and 0.01875IU/mL demonstrated that the T. pallidum microsieve assay is able to distinguish disease-specific IgG signal from background signal at similar, and possibly lower, levels than the corresponding ELISA. The T. pallidum microsieve assay prototype also differentiated positive clinical serum samples from negative donor samples, and the results were in good agreement with ELISA (R(2)=0.9046). These feasibility studies demonstrate the potential for utilizing microsieves, along with a reagent wicking device, as a simple diagnostic immunoassay platform.
authors.affiliation: ZEUS Scientific Inc., 200 Evans Way, Branchburg, NJ 08876, USA. Electronic address: dzweitzig@zeusscientific.com.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Zweitzig
authors.affiliation: Department of Research & Development, VyCAP, Hallenweg 23, 7522 NH Enschede, The Netherlands.
authors.forename: Arjan G
authors.initials: AG
authors.lastname: Tibbe
authors.affiliation: Aquamarijn Micro Filtration B.V., Ijsselkade 7, 7201 HB Zutphen, The Netherlands.
authors.forename: Ai T
authors.initials: AT
authors.lastname: Nguyen
authors.affiliation: Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB Wageningen, The Netherlands.
authors.forename: Cees J M
authors.initials: CJ
authors.lastname: van Rijn
authors.affiliation: ZEUS Scientific Inc., 200 Evans Way, Branchburg, NJ 08876, USA.
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Kopnitsky
authors.affiliation: ZEUS Scientific Inc., 200 Evans Way, Branchburg, NJ 08876, USA.
authors.forename: Kathleen
authors.initials: K
authors.lastname: Cichonski
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute, University of Twente, Hallenweg 23, 7522 NH Enschede, The Netherlands.
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D025065:Borrelia burgdorferi / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008193:Lyme Disease / Q000175:diagnosis*; D012327:RNA Virus Infections / Q000175:diagnosis*; D012015:Reference Standards; D012409:Rubella / Q000276:immunology*; D012680:Sensitivity and Specificity; D012825:Silicon; D013587:Syphilis / Q000175:diagnosis*; D014210:Treponema pallidum / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D000914:Antibodies, Viral; C040901:rubella antibodies; D012825:Silicon
keywords: Diagnosis; Disease; Immunoassay; Infectious; Microsieve; Serology