Advances in membrane receptor screening and analysis

Abstract

During the last decade there has been significant progress in the development of analytical techniques for the screening of ligand binding to membranes and membrane receptors. This review focuses on developments using label-free assays that facilitate ligand-membrane-receptor screening without the need for chemical-, biological- or radiological-labelled reagents. These assays include acoustic, optical surface plasmon resonance biosensing, sedimentation (analytical ultracentrifugation), chromatographic assays, isothermal titration calorimetry and differential scanning calorimetry. The merits and applications of cell-based screening systems and of different model membrane systems, including planar supported lipid layers, bead-supported membranes and lipid micro-arrays, are discussed. Recent advances involving more established techniques including intrinsic fluorescence, FRET spectroscopy, scintillation proximity assays and automated patch clamping are presented along with applications to peripheral membrane proteins, ion channels and G protein-coupled receptors. Novel high-throughput assays for determination of drug- and protein-partitioning in membranes are also highlighted. To aid the experimenter, a brief synopsis of the techniques commonly employed to purify and reconstitute membranes and membrane receptors is included.

Nanoliter contact angle probes tumor angiogenic ligand–receptor protein interactions

“…Solution phase assays, such as micro-calorimetric (Falconer et al, 2010), are affected by limitations related to the use of large amounts of highly purified proteins (milliliters of pro-tein solutions at M concentrations, at least) and to the different conformation that protein can assume in solution with respect to cellular environments (Cooper, 2004). On the contrary, solid phase assays performed on intact cells, cell membrane preparations or with immobilization of the receptors in "cell-like" environments have been used to reveal protein-protein interactions and to study their kinetics.…”

Melting and interdigitation of microstructured solid supported membranes quantified by imaging ellipsometry

“…[9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26] However, little is known about the impact of the solid support on physical properties of lipid bilayers attached to surfaces such as the main phase transition temperature or lateral mobility of the lipids. It is expected that phase transitions are strongly affected if not entirely suppressed by the various different supports ranging from glass, gold, silicon, mica to all kinds of functionalized surfaces.…”

Survey of the year 2004 commercial optical biosensor literature

“…cell adhesion [64], innate immunity [3], CO 2 assimilation in metabolic pathways [30], extracellular matrix degradation [2] and membrane construction [17,44,85]) as well as the binding properties of retrocyclins [14], apotosis regulators [42], antimicrobial peptides [23], glycosphingolipids [97], barley -amylase/subtilisin inhibitor [66], amelogenin [67], lysenin [45], lectins [65], endoxylases [40], connexins [21], selectins [25], phosphoinositides [71], carbohydrates [20,70,72], HIV proteins [48,91], tumor necrosis factor receptor superfamily [95] and the insulin-like growth factor system [58]. Other reviews highlight the integration of optical biosensors with complementary technologies in areas of applied research, including drug development and chemical genetics [1,50,73,83,88], plant proteomics [33], gene and protein expression [29,35,86], antibody optimization [5,46,62], food industry [4,47,54,61,79], bioprocessing and manufacturing [12,…”