Multiplexing ligand-receptor binding measurements by chemically patterning microfluidic channels.
A method has been designed for patterning supported phospholipid bilayers (SLBs) on planar substrates and inside microfluidic channels. To do this, bovine serum albumin (BSA) monolayers were formed via adsorption at the liquid/solid interface. Next, this interfacial protein film was selectively patterned by using deep UV lithography. Subsequently, SLBs could be deposited in the patterned locations by vesicle fusion. By cycling through this process several times, spatially addressed bilayer arrays could be formed with intervening protein molecules serving as two-dimensional corrals. By employing this method, phospholipid bilayers containing various concentrations of ganglioside GM1 were addressed along the length of individual microfluidic channels. Therefore, the binding of GM1 with pentameric cholera toxin B (CTB) subunits could be probed. A seven-channel microfluidic device was fabricated for this purpose. Each channel was simultaneously patterned with four chemically distinct SLBs containing 0, 0.2, 0.5, and 2.0 mol % GM1, respectively. Varying concentrations of CTB were then introduced into each of the channels. With the use of total internal reflection fluorescence microscopy, it was possible to simultaneously abstract multiple equilibrium dissociation constants as a function of ligand density for the CTB-GM1 system in a single shot.
Department of Chemistry, Texas A&M University, College Station, Texas 77843, USA.
Jinjun
J
Shi
Tinglu
T
Yang
Paul S
PS
Cremer
D000818:Animals; D002417:Cattle; D002772:Cholera Toxin / Q000378:metabolism; D005677:G(M1) Ganglioside / Q000378:metabolism; D008051:Lipid Bilayers; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D054802:Molecular Imprinting / Q000379:methods*; D010743:Phospholipids; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000295:instrumentation / Q000379:methods*; D012710:Serum Albumin, Bovine
D008051:Lipid Bilayers; D010743:Phospholipids; D012710:Serum Albumin, Bovine; D005677:G(M1) Ganglioside; D002772:Cholera Toxin