Development of a Micro Total Analysis System Incorporating Chemiluminescence Detection and Application to Detection of Cancer Markers

Abstract

We developed a micro total analysis system (mu-TAS) incorporating chemiluminescence detection, in which the chemiluminescence reaction of isoluminol isothiocyanato (ILITC) (as a chemiluminescence reagent for labeling)-microperoxidase (as a catalyst)-hydrogen peroxide (as an oxidant) was adopted. The analysis system performed the following three processes on a microchip: immune reaction for high selectivity, electrophoresis for formation and transportation of the sample plug, and chemiluminescence detection for high sensitivity. The three processes were compactly integrated onto the microchip to give the mu-TAS. The microchip contained two microchannels that crossed at an intersection, while the ends of the microchannels accessed four reservoirs. As the first process, the immune reaction was performed using an antibody-immobilized glass bead. The glass bead was placed in one of the reservoirs along with antigen (analyte) and a known amount of ILITC-labeled antigen to set up a competitive immune reaction. For electrophoresis, as the second process, the reactant after the immune reaction was fed electrophoretically into the intersection resulting in a sample plug. The sample plug was then moved into another reservoir containing hydrogen peroxide solution. At this point, chemiluminescence detection was performed as the third process: the labeled antigen mixed with the hydrogen peroxide and the catalyst included in the migration buffer to produce chemiluminescence. Chemiluminescence was detected by a photomultiplier tube located under the reservoir. The mu-TAS described here was capable of determining, with high selectivity and sensitivity, human serum albumin or immunosuppressive acidic protein as a cancer marker in human serum.

Capillary electrophoresis of proteins 2003–2005

“…For hemoglobin, a detection limit of 1.2610 27 M (0.6 fmol) was achieved. The chemiluminescent detector was also modified for CE on chip [34]. Immunoaffinity interactions can also be used for specific labeling of analyzed proteins.…”

“…For a chemiluminescence detection of proteins, a derivatization with isoluminol thiocyanate has been introduced [34].…”

“…A number of CE steps and components that are used in fused-silica capillaries have been transferred and successfully tested on a microchip including preconcentration on porous silica membrane [22,23], automated derivatization [29], microreactor with immobilized proteases and other proteins [181,182], UV detector [183,184], chemiluminescence detector [34,185], multiwavelength LIF detector [186], UV LIF detector [98], wall coatings [82,85], affinity CE [129], CIEF [80], and sheathless electrospray for MS [113,114].…”

Chemiluminescence immunoassay

“…Micro total analytical system (l-TAS) incorporating immunoassay and chemiluminescence detection [112].…”

Unconventional detection methods for microfluidic devices

“…As a continuation of their work, the same group analyzed dansylated amino acids with a bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H 2 O 2 system in their detector design [80]. Tsukagoshi et al [81] followed with a setup that used modified versions of a commercially available instrument and CL detector. They analyzed HSA and immunosuppressive acidic protein (a cancer marker) in human serum.…”