Online Preconcentration by Transient Isotachophoresis in Linear Polymer on a Poly(methyl methacrylate) Microchip for Separation of Human Serum Albumin Immunoassay Mixtures

Abstract

Online preconcentration of human serum albumin (HSA) and its immunocomplex with a monoclonal antibody by on-chip transient isotachophoresis is reported. An 800-fold signal enhancement was achieved following the preconcentration on standard cross-channel microchips made of poly (methyl methacrylate). Sample injection, preconcentration, and separation were done continuously and controlled solely by a sequential voltage switching program. The preconcentration was followed by on-chip nondenaturing gel electrophoresis in methylcellulose solution. The method was applied to microchip electrophoresis immunoassay of HSA. Baseline separation of HSA and its immunocomplex was achieved in 25 s in the first 1 cm of the microchannel. In a direct immunoassay, the minimum detectable concentration of fluorescent labeled HSA by laser-induced fluorescence detection was 7.5 pM.

Nano- and Microbiodevices for High-Performance Separation of Biomolecules

“…7 and 8 [8][9][10][11][12][13][14][15][16][17][18][19][20][21]. Nanobiodevices are applied to diagnosis of hypertension by the fast analysis of SNPs (single nucleotide polymorphisims) of hypertension related genes and diagnosis of lung cancer through the detection of deletion of the specific sequence for the lung cancer related genes.…”

Fabrication, modification, and application of poly(methyl methacrylate) microfluidic chips

“…Online preconcentration of HSA and its immunocomplex with a mAb by on-chip transient ITP with LIF detection has been reported by Mohamadi et al [98]. Bai et al [99] developed a novel surface treatment method using PEI to enhance antibody binding on the PMMA microfluidic immunoassay device.…”

Nanobiodevice-based Single Biomolecule Analysis, Single-Cell Analysis, and in vivo Imaging for Cancer Diagnosis, Cancer Theranostics, and iPS Cell-based Regenerative Medicine

“…[16][17][18] The research efforts in my laboratory have been focused on the development of novel nanobiodevices for biomedical applications. [1][2][3][4][5][6][7] In this review article, we mainly overview our research activities, including nanobiodevices for single-cell analysis, [20][21][22][23][24][25] single biomolecule analysis, 19,[26][27][28][29]41,46,50,[54][55][56][57][58] biomarker detections, 23,[30][31][32][33]39,[47][48][49] cancer theranostics, and iPS cell in vivo imaging. [20][21][22][23][24][25][34][35][36][37][38]…”