Use of a mixture of n-dodecyl-beta-D-maltoside and sodium dodecyl sulfate in poly(dimethylsiloxane) microchips to suppress adhesion and promote separation of proteins.
Dynamic modification of poly(dimethylsiloxane) channels using a mixture of n-dodecyl-beta-D-maltoside (DDM) and sodium dodecyl sulfate (SDS) is able to suppress analyte adsorption and control electroosmotic flow (EOF). In this mixed surfactant system, the nonionic surfactant DDM functions as a surface blocking reagent, whereas the anionic surfactant SDS introduces negative charges to the channel walls. Changing the DDM/SDS mixing ratio tunes the surface charge density and the strength of EOF. Using 0.1% (w/v) DDM and 0.03% (w/v) SDS, Alexa Fluor 647 labeled streptavidin can be analyzed according to the charges added by the fluorophores. Protein molecules with different numbers of fluorophores are well resolved. DDM and SDS also form negatively charged mixed micelles, which act as a separation medium. The low critical micellar concentration of DDM/SDS mixed micelles also allows the use of SDS at a nondenaturing concentration, which enables the analysis of proteins in their native state. The immunocomplex between a membrane protein, beta2 adrenergic receptor, and anti-FLAG antibody has been fully separated using 0.1% (w/v) DDM and 0.03% (w/v) SDS. We have also analyzed the composition of light-harvesting protein-chromophore complexes in cyanobacteria.
Huang
Bo
B
Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
Kim
Samuel
S
Wu
Hongkai
H
Zare
Richard N
RN
D000458:Cyanobacteria / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D046209:Electrophoresis, Microchip; D008320:Maltose / Q000031:analogs & derivatives* / Q000737:chemistry; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*; D012828:Silicones / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*
D004129:Dimethylpolysiloxanes; D011506:Proteins; D012828:Silicones; C411362:dodecyl maltopyranoside; D012967:Sodium Dodecyl Sulfate; C013830:baysilon; D008320:Maltose