Described herein are methods of separating lipid nanoparticles (LNPs) according to their isoelectic points, the methods comprising applying a separating voltage to a separation matrix comprising carrier ampholytes and the LNPs for a sufficient time to separate the LNPs according to their isoelectic points.

Some embodiments described herein relate to a method that includes separating an analyte-containing sample via electrophoresis in a capillary. The capillary is loaded with a chemiluminescence agent, such as luminol, that is configured to react with the analyte (e.g., HRP-conjugated proteins) to produce a signal indicative of a concentration and/or quantity of analyte at each location along the length of the capillary. A first image of the capillary containing the analytes and the chemiluminescence agent is captured over a first period of time. A second image of the capillary containing the analytes and the chemiluminescence agent is captured over a second, longer, period of time. A concentration and/or quantity of a first population of analytes at a first location is determined using the first image, and a concentration and/or quantity of a second population of analytes at a second location is determined using the second image.

Some embodiments described herein relate to a method that includes separating an analyte-containing sample via electrophoresis in a capillary. The capillary is loaded with a chemiluminescence agent, such as luminol, that is configured to react with the analyte (e.g., HRP-conjugated proteins) to produce a signal indicative of a concentration and/or quantity of analyte at each location along the length of the capillary. A first image of the capillary containing the analytes and the chemiluminescence agent is captured over a first period of time. A second image of the capillary containing the analytes and the chemiluminescence agent is captured over a second, longer, period of time. A concentration and/or quantity of a first population of analytes at a first location is determined using the first image, and a concentration and/or quantity of a second population of analytes at a second location is determined using the second image.

The disclosure provides a method of separating a protein product, e.g., from a contaminant in a mixture, comprising contacting the mixture with a polishing chromatography matrix. In some embodiments, the polishing chromatography matrix is used together with a retained gradient. In certain embodiments, the retained gradient comprises a pH gradient. In certain embodiments, the mixture comprises a monoclonal antibody product of a prior protein A chromatography.

The disclosure provides a method of separating a protein product, e.g., from a contaminant in a mixture, comprising contacting the mixture with a polishing chromatography matrix. In some embodiments, the polishing chromatography matrix is used together with a retained gradient. In certain embodiments, the retained gradient comprises a pH gradient. In certain embodiments, the mixture comprises a monoclonal antibody product of a prior protein A chromatography.

Microfluidic devices and nanobiosensors comprising a magnetic nanoparticle attached to a reporter molecule via a release unit for microfluidic-based detection of a target analyte in a biological sample. The nanobiosensors can be magnetically manipulated or guided through the microfluidics channels for incubation with the biological sample, concentration of the nanobiosensors, and detection of target analytes, without having to pump the entire initial sample through a microfluidic channel of the microfluidic device. The magnetic nanoparticles are separated from the reporter molecules before detection and can be re-used.

Microfluidic devices and nanobiosensors comprising a magnetic nanoparticle attached to a reporter molecule via a release unit for microfluidic-based detection of a target analyte in a biological sample. The nanobiosensors can be magnetically manipulated or guided through the microfluidics channels for incubation with the biological sample, concentration of the nanobiosensors, and detection of target analytes, without having to pump the entire initial sample through a microfluidic channel of the microfluidic device. The magnetic nanoparticles are separated from the reporter molecules before detection and can be re-used.

Embodiments of the invention are directed to a method of separating and encapsulating an analyte on a microfluidic device in order to extract the analyte. A microfluidic device is provided having a main microchannel and a set of one or more auxiliary microchannels, each branching to the main microchannel at respective junctions therewith. A mixture is introduced as a single phase in the main microchannel in order to electrokinetically separate an analyte from the introduced mixture, and in order to confine the separated analyte in a microchannel portion of the main microchannel. The microchannel portion adjoins one of the junctions. One or more encapsulating volumes of an encapsulating phase are injected in the main microchannel via one or more of the auxiliary microchannels. The encapsulating phase is immiscible with said single phase. The encapsulated analyte is extracted from the main microchannel via one or more of the auxiliary microchannels.

Embodiments of the invention are directed to a method of separating and encapsulating an analyte on a microfluidic device in order to extract the analyte. A microfluidic device is provided having a main microchannel and a set of one or more auxiliary microchannels, each branching to the main microchannel at respective junctions therewith. A mixture is introduced as a single phase in the main microchannel in order to electrokinetically separate an analyte from the introduced mixture, and in order to confine the separated analyte in a microchannel portion of the main microchannel. The microchannel portion adjoins one of the junctions. One or more encapsulating volumes of an encapsulating phase are injected in the main microchannel via one or more of the auxiliary microchannels. The encapsulating phase is immiscible with said single phase. The encapsulated analyte is extracted from the main microchannel via one or more of the auxiliary microchannels.

The present disclosure provides a method and apparatus for improvements of sample throughput in proteome analysis by mass spectrometry, by combining multiple non-overlapping isoelectric focusing separations. The method for performing an analysis of a plurality of protein samples, comprises: (a) Adding a proteolytic enzyme of a given specificity to a first protein sample to digest proteins to peptides; (b) Separating the peptides obtained in step (a) by isoelectric focusing; (c) Collecting those peptides which have their isoelectric point value within a first isoelectric point range; (d) Adding a proteolytic enzyme of a given specificity to a second protein sample to digest proteins to peptides; (e) Separating the peptides obtained in step (d) by isoelectric focusing; (f) Collecting those peptides which have their isoelectric point value within a second isoelectric point range, where said second isoelectric point range is different and non-overlapping compared to said first isoelectric point range; (g) Combining the peptides collected in steps (c) and (f) into a single sample and subjecting said sample to mass spectrometry analysis; (h) Deconvoluting signals/data obtained from the mass spectrometry analysis by calculating the isoelectric point of each peptide, and assigning a peptide to the first protein sample if its isoelectric point value matches the isoelectric point range selected in step (c) or to the second protein sample if its isoelectric point value matches the isoelectric point range selected in step (f); and (i) Obtaining quantitative information for proteins of each sample according to magnitude of the signal obtained from each peptide.