Carboxymethyl cellulose film as a substrate for microarray fabrication.
Magnetic beads (MB) are widely used for quick and highly sensitive signal detection in microarray-based assays. However, this technique imposes stringent requirements for smoothness and adhesive properties of the surface, which most common substrates do not satisfy. We report here a new type of substrate for microarrays with a low adhesion to MB-thermally cross-linked carboxymethyl cellulose (CMC) film. This substrate can be readily fabricated on a conventional glass slide. A highly cross-linked CMC film (∼1 cross-link per monomer unit) possesses a surface smooth on a nanometer scale and a low adhesion to protein-coated MB, which partly originates from electrostatic repulsion of MB from negatively charged CMC surface. The efficiency of the CMC substrate is demonstrated hereby in fabrication of microarrays for the detection of three bacterial toxins: cholera toxin, staphylococcal enterotoxin A, and toxic shock syndrome toxin. The assay employing a primary antibodies arrayed on a CMC surface and detection of the bound bacterial toxins with a biotinylated secondary antibodies and streptavidin-coated MB resulted in a limits of detection as low as 0.1 ng/mL. The CMC-based microarrays demonstrated very high storage stability; their activity did not change after one year storage at room temperature.
Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences , Pushchino, Moscow Region, Russia 142290.
Yuri M
YM
Shlyapnikov
Elena A
EA
Shlyapnikova
Victor N
VN
Morozov
D000269:Adhesives / Q000737:chemistry / Q000378:metabolism; D002266:Carboxymethylcellulose Sodium / Q000737:chemistry* / Q000378:metabolism; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D013379:Substrate Specificity; D013499:Surface Properties
D000269:Adhesives; D058185:Magnetite Nanoparticles; D002266:Carboxymethylcellulose Sodium