The present invention relates to the unexpected discovery of methods and devices that can be used for high-throughput precise quantification, detection and/or temporal profiling of cellular secretions. In various embodiments, the methods of the invention allow for high-throughput absolute detection of secretions of cells, identification of the nature of the secreted molecules, and/or the nature of the secreting cells. Further, the present invention includes a device combining microfluidics and antibody printing, wherein the device can be used to detect protein secretion signature of cells in a high-throughput manner. Further, the present invention includes compositions comprising molecules that can be used to reduce cell death and to implement cell-less therapies. Further, the present invention includes a method for training an algorithm to predict temporal profile of cellular secretion.

The present invention relates to the unexpected discovery of methods and devices that can be used for high-throughput precise quantification, detection and/or temporal profiling of cellular secretions. In various embodiments, the methods of the invention allow for high-throughput absolute detection of secretions of cells, identification of the nature of the secreted molecules, and/or the nature of the secreting cells. Further, the present invention includes a device combining microfluidics and antibody printing, wherein the device can be used to detect protein secretion signature of cells in a high-throughput manner. Further, the present invention includes compositions comprising molecules that can be used to reduce cell death and to implement cell-less therapies. Further, the present invention includes a method for training an algorithm to predict temporal profile of cellular secretion.

Various embodiments of an apparatus for measuring binding kinetics of an interaction of an analyte material present in a fluid sample are disclosed. The apparatus includes a sensing resonator having at least one binding site for the analyte material; actuation circuitry adapted to drive the sensing resonator into an oscillating motion; measurement circuitry coupled to the sensing resonator and adapted to measure an output signal of the sensing resonator representing resonance characteristics of the oscillating motion of the sensing resonator; and a controller coupled to the actuation and measurement circuitry, wherein the controller is adapted to detect an individual binding event between the at least one binding site and a molecule of the analyte material.

Various embodiments of an apparatus for measuring binding kinetics of an interaction of an analyte material present in a fluid sample are disclosed. The apparatus includes a sensing resonator having at least one binding site for the analyte material; actuation circuitry adapted to drive the sensing resonator into an oscillating motion; measurement circuitry coupled to the sensing resonator and adapted to measure an output signal of the sensing resonator representing resonance characteristics of the oscillating motion of the sensing resonator; and a controller coupled to the actuation and measurement circuitry, wherein the controller is adapted to detect an individual binding event between the at least one binding site and a molecule of the analyte material.

What has been developed is an improved method of determining the binding constants between two molecules that requires significantly fewer materials and potentially less time (in the case of a whole cell analysis less time to grow cell cultures as fewer cells are required in the same analysis) to undertake. The method involves utilizing an NSB measurement in preferably an n-curve analysis in order to determine the K.sub.d and/or R.sub.t without having to complete actual measurements to determine the lower knee of the curve(s) in the n-curve analysis. Preparing the experiment utilizing the additional samples allows an experiment represented in a binding curve having an upper and a lower knee to no longer be required to run through the lower knee to a point of completion.

What has been developed is an improved method of determining the binding constants between two molecules that requires significantly fewer materials and potentially less time (in the case of a whole cell analysis less time to grow cell cultures as fewer cells are required in the same analysis) to undertake. The method involves utilizing an NSB measurement in preferably an n-curve analysis in order to determine the K.sub.d and/or R.sub.t without having to complete actual measurements to determine the lower knee of the curve(s) in the n-curve analysis. Preparing the experiment utilizing the additional samples allows an experiment represented in a binding curve having an upper and a lower knee to no longer be required to run through the lower knee to a point of completion.

A system and method for evaluation of an interaction between an analyte in a fluid sample and a ligand immobilized on a sensor surface of a biosensor is provided. In one example, the system includes a plurality of needles, each being arranged to inject a fluid sample to one of sensor surfaces or detection spots. A plurality of fluid samples, each containing known concentrations of analyte, is provided. The plurality of fluid samples may be divided into at least two groups, each group having a number of fluid samples corresponding to the number of needles. The system and method is configured to perform the injections without intermediate regeneration or renewal of the immobilized ligand. Software for performing the steps of the method and a computer readable medium for storing the software are also provided.

A system and method for evaluation of an interaction between an analyte in a fluid sample and a ligand immobilized on a sensor surface of a biosensor is provided. In one example, the system includes a plurality of needles, each being arranged to inject a fluid sample to one of sensor surfaces or detection spots. A plurality of fluid samples, each containing known concentrations of analyte, is provided. The plurality of fluid samples may be divided into at least two groups, each group having a number of fluid samples corresponding to the number of needles. The system and method is configured to perform the injections without intermediate regeneration or renewal of the immobilized ligand. Software for performing the steps of the method and a computer readable medium for storing the software are also provided.

The present disclosure provides, inter alia, genetically encoded recombinant peptide biosensors comprising analyte-binding framework portions and signaling portions, wherein the signaling portions are present within the framework portions at sites or amino acid positions that undergo a conformational change upon interaction of the framework portion with an analyte.

The present disclosure provides, inter alia, genetically encoded recombinant peptide biosensors comprising analyte-binding framework portions and signaling portions, wherein the signaling portions are present within the framework portions at sites or amino acid positions that undergo a conformational change upon interaction of the framework portion with an analyte.