title: Programmable bio-nano-chip systems for serum CA125 quantification: toward ovarian cancer diagnostics at the point-of-care.
abstract: Point-of-care (POC) implementation of early detection and screening methodologies for ovarian cancer may enable improved survival rates through early intervention. Current laboratory-confined immunoanalyzers have long turnaround times and are often incompatible with multiplexing and POC implementation. Rapid, sensitive, and multiplexable POC diagnostic platforms compatible with promising early detection approaches for ovarian cancer are needed. To this end, we report the adaptation of the programmable bio-nano-chip (p-BNC), an integrated, microfluidic, and modular (programmable) platform for CA125 serum quantitation, a biomarker prominently implicated in multimodal and multimarker screening approaches. In the p-BNCs, CA125 from diseased sera (Bio) is sequestered and assessed with a fluorescence-based sandwich immunoassay, completed in the nano-nets (Nano) of sensitized agarose microbeads localized in individually addressable wells (Chip), housed in a microfluidic module, capable of integrating multiple sample, reagent and biowaste processing, and handling steps. Antibody pairs that bind to distinct epitopes on CA125 were screened. To permit efficient biomarker sequestration in a three-dimensional microfluidic environment, the p-BNC operating variables (incubation times, flow rates, and reagent concentrations) were tuned to deliver optimal analytical performance under 45 minutes. With short analysis times, competitive analytical performance (inter- and intra-assay precision of 1.2% and 1.9% and limit of detection of 1.0 U/mL) was achieved on this minisensor ensemble. Furthermore, validation with sera of patients with ovarian cancer (n = 20) showed excellent correlation (R(2) = 0.97) with gold-standard ELISA. Building on the integration capabilities of novel microfluidic systems programmed for ovarian cancer, the rapid, precise, and sensitive miniaturized p-BNC system shows strong promise for ovarian cancer diagnostics.
authors.affiliation: Departments of Bioengineering and Chemistry, Rice University, Houston, TX 77005, USA.
authors.forename: Archana
authors.initials: A
authors.lastname: Raamanathan
authors.forename: Glennon W
authors.initials: GW
authors.lastname: Simmons
authors.forename: Nicolaos
authors.initials: N
authors.lastname: Christodoulides
authors.forename: Pierre N
authors.initials: PN
authors.lastname: Floriano
authors.forename: Wieslaw B
authors.initials: WB
authors.lastname: Furmaga
authors.forename: Spencer W
authors.initials: SW
authors.lastname: Redding
authors.forename: Karen H
authors.initials: KH
authors.lastname: Lu
authors.forename: Robert C
authors.initials: RC
authors.lastname: Bast
authors.forename: John T
authors.initials: JT
authors.lastname: McDevitt
mesh_terms: D001774:Blood Chemical Analysis / Q000295:instrumentation / Q000379:methods / Q000592:standards; D018394:CA-125 Antigen / Q000032:analysis* / Q000097:blood; D002138:Calibration; D002277:Carcinoma / Q000097:blood / Q000175:diagnosis*; D003201:Computers / Q000592:standards; D055088:Early Detection of Cancer / Q000295:instrumentation / Q000379:methods / Q000592:standards / Q000639:trends; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods / Q000592:standards; D005260:Female; D006801:Humans; D056656:Lab-On-A-Chip Devices / Q000592:standards; D046208:Microchip Analytical Procedures / Q000379:methods* / Q000592:standards; D008954:Models, Biological; D009994:Osmolar Concentration; D010051:Ovarian Neoplasms / Q000097:blood / Q000175:diagnosis*; D019095:Point-of-Care Systems / Q000639:trends
chemical_list: D018394:CA-125 Antigen