Biomedical subjects

Tinglu Yang

Publications and source records attributed to Tinglu Yang.

3 recordsLinked to original sources

PubMed2003

Investigations of bivalent antibody binding on fluid-supported phospholipid membranes: the effect of hapten density.

Investigations of ligand-receptor binding between bivalent antibodies and membrane-bound ligands are presented. The purpose of these studies was to explore binding as a function of hapten density in a two-dimensionally fluid environment. A novel microfluidic strategy in conjunction with total internal reflection fluorescence microscopy was designed to achieve this. The method allowed binding curves to be acquired with excellent signal-to-noise ratios while using only minute quantities of protein solution. The specific system investigated was the interaction between anti-DNP antibodies and phospholipid membranes containing DNP-conjugated lipids. Binding curves for ligand densities ranging from 0.1 to 5.0 mol % were obtained. Two individual dissociation constants could be extracted from the data corresponding to the two sequential binding events. The first dissociation constant, K(D1), was 2.46 x 10(-)(5) M, while the second was K(D2) = 1.37 x 10(-)(8) mol/m(2). This corresponded to a positively cooperative binding effect with an entropic difference between the two events of 62.3 +/- 2.7 J/(mol.K). Furthermore, the percentage of monovalently and bivalently bound protein was determined at each ligand density.

2,4-Dinitrophenol↗

PubMed2002

A microfluidic device with a linear temperature gradient for parallel and combinatorial measurements.

Methods for obtaining combinatorial and array-based data as a function of temperature are needed in the chemical and biological sciences. It is presently quite difficult to employ temperature as a variable using standard wellplate formats simply because it is very inconvenient to keep each well at a distinct temperature. In microfluidics, however, the situation is very different due to the short length scales involved. In this article, it is shown how a simple linear temperature gradient can be generated across dozens of parallel microfluidic channels simultaneously. This result is exploited to rapidly obtain activation energies from catalytic reactions, melting point transitions from lipid membranes, and fluorescence quantum yield curves from semiconductor nanocrystal probes as a function of temperature. The methods developed here could quite easily be extended to protein crystallization, phase diagram measurements, chemical reaction optimization, or multivariable experiments.

Alkaline Phosphatase↗

PubMed2002

Design and characterization of immobilized enzymes in microfluidic systems.

Herein we report the fabrication, characterization, and use of total analytical microsystems containing surface-immobilized enzymes. Streptavidin-conjugated alkaline phosphatase was linked to biotinylated phospholipid bilayers coated inside poly(dimethylsiloxane) microchannels and borosilicate microcapillary tubes. Rapid determination of enzyme kinetics at many different substrate concentrations was made possible by carrying out laminar flow-controlled dilution on-chip. This allowed Lineweaver-Burk analysis to be performed from a single experiment with all the data collected simultaneously. The results revealed an enzyme turnover number of 51.1 +/- 3.2 s(-1) for this heterogeneous system. Furthermore, the same enzyme immobilization strategy was extended to demonstrate that multiple chemical reactions could be performed in sequence by immobilizing various enzymes in series. Specifically, the presence of glucose was detected by two coupled steps employing immobilized avidinD-conjugated glucose oxidase and streptavidin-conjugated horseradish peroxidase.

Bacterial Proteins↗