Analysis of inflammatory biomarkers from tissue biopsies by chip‐based immunoaffinity CE

Abstract

To aid in the biochemical analysis of human skin biopsies, a semiautomatic chip-based CE system has been developed for measuring inflammatory biomarkers in microdissected areas of the biopsy. Following solubilization of the dissected tissue, the desired biomarkers were isolated by immunoaffinity capture using a panel of 12 antibodies, immobilized on a disposable glass fiber disk, within the extraction port of the chip. The captured analytes were labeled with a 635 nm light-emitting laser dye and electroeluted into the separation channel. Electrophoretic separation of all of the analytes was achieved in 2.2 min with quantification of each peak being performed by online LIF detection and integration of each peak area. Comparison of the results obtained from the chip-based system to those obtained using commercially available high-sensitivity immunoassays demonstrated that the chipbased assay provides a fast, accurate procedure for studying the concentrations of inflammatory biomarkers in complex biological materials. The degree of accuracy and precision achieved by the chip-based CE is comparable to conventional immunoassays and the system is capable of analyzing circa six samples per hour. With the ever-expanding array of antibodies that are commercially available, this chip-based system can be applied to a wide variety of different biomedical analyses.

Immunoaffinity capillary electrophoresis: A new versatile tool for determining protein biomarkers in inflammatory processes

“…Use of the IACE technology with antibodies or antibody fragments has been reported independently since the early 1990s by the Guzman and Phillips groups [32,33,[100][101][102][103][104][105][106] (T. M. Phillips, paper in preparation). Figure 4 shows a typical electropherogram representing the quantification of a number of chemokines present in human skin biopsy extracts.…”

Immunoaffinity Chromatography: A Review

“…In recent years, IAC has been integrated with other analytical methods such as CE and MS. CE immunoassays (where antibodies are immobilized in CE capillaries) are utilized because they are easily automated, require small amounts of sample and reagents and still maintain a good detection limit, and offer relatively fast separation (Moser & Hage, 2008;Phillips and Wellner, 2007). …”

Clinically relevant advances in on‐chip affinity‐based electrophoresis and electrochromatography

“…In a later work, Phillips and Wellner [77] developed a microdevice to automate sample incubation, affinity-based analyte isolation, laser dye labeling, and analyte elution with pressure-driven and electrokinetic flow. The device was utilized to quantify 12 inflammatory biomarkers from human skin biopsies.…”