Abstract: The present disclosure provides compositions and methods for high throughput Gain of Function (GOF) sorting to discover and develop novel and highly selective candidate drug molecules. High throughput GOF sorting includes: mutating a single residue in a receptor and/or ligand; measuring the affinity and functional activity of the resulting ligand target-ligand interaction; and carrying out multiple rounds of mutation and measurement to determine which residues provide key interaction points underlying the functional activity of a ligand target-ligand interaction. Further, the present disclosure provides methods and compositions for high throughput and high precision GOF sorting, such that large numbers of mutations can be generated and screened rapidly, and GOF compounds can be identified and isolated.

Abstract: The present disclosure provides systems for multiplexed multitarget screening of cell populations having one or more wild type or mutated ligand targets and measuring cell responses to ligands using high throughput screening techniques, including flow cytometry (FCM). The method includes the steps of: 1) developing cell populations to be screened; 2) staining cell populations using one or more fluorochromes to yield a distinct excitation/emission signature for each cell population; 3) combining labelled cell populations into a single mixed suspension; 4) analyzing populations to resolve them on the basis of their unique signature; and 5) resolving individual populations and deconvoluting data to extract meaningful information about populations.

Abstract: An apparatus and method for real-time measurement of a cellular response of a test compound or series of test compounds (303) on a flowing suspension of cells (349), in which a homogeneous suspension of each member of a series of cell types (349) is combined with a concentration of a test compound (303), directed through a detection zone (355), and a cellular response of the living cells is measured in real time as the cells in the test mixture are flowing through the detection zone (355). The apparatus may be used in automated screening of libraries of compounds, and is capable of real-time variation of concentrations of test and standard compounds and generation of dose/response profiles within a short time span.

Abstract: An apparatus and method for real-time measurement of a cellular response of a test compound or series of test compounds (303) on a flowing suspension of cells (349), in which a homogeneous suspension of each member of a series of cell types (349) is combined with a concentration of a test compound (303), directed through a detection zone (355), and a cellular response of the living cells is measured in real time as the cells in the test mixture are flowing through the detection zone (355). The apparatus may be used in automated screening of libraries of compounds, and is capable of real-time variation of concentrations of test and standard compounds and generation of dose/response profiles within a short time span.

Abstract: The improvement of thin tunnel oxides used in EEPROM and FLASH tecnologies using post-oxidation annealing in nitrogen causes defects in subsequent oxide films. These are manifested by oxide thinning at the bird's beak and result in high gate leakage. As the time and temperature to the post-oxidation annealing are increased for improved tunnel oxide performance, the number of defects increases rapidly. A method of realizing the improved tunnel oxide Q.sub.BD using higher post-oxidation time and temperature annealing while at the same time not degrading the quality of subsequent gate oxides is shown. The use of sacrificial oxidation and strip just prior to the transistor gate oxidation is described. This process removes the additional nitride which exists at the field edges, leading to the oxide thinning. As a result, improved tunnel oxide integrity can be achieved without degradation of high and low voltage transistors.