Abstract:
A method of generating a protein with an improved functional property, the method comprising: (a) identifying at least one Target amino acid Residue in a first protein, wherein said Target amino acid Residue is associated with said functional property; (b) comparing at least one homologous second protein from the same or a different phylogenetic branch as the first protein with the first protein and identifying at least one Variant amino acid Residue between the first protein and the second protein; (c) selecting at least one Candidate amino acid Residue from the Variant amino acid Residue identified in (b) on the basis of said Candidate amino acid Residue affecting said Target amino acid Residue with respect to said functional property; (d) forming at least one Candidate Mutant protein in silico or producing at least one Candidate Mutant protein in vitro in which said at least one Candidate amino acid Residue from the second protein substitutes a corresponding residue in the first protein; and (e) screening said at least one Candidate Mutant protein produced in (d) to identify a protein having said improved functional property.
Abstract:
Described are methods for determination of peptide specific parameter(s) of a mixture comprising a target peptide and a related impurity (or impurities) to be used in a simulation model of chromatographic separation using mathematical model(s). Also described are chromatographic simulation methods using above determined parameters, as well as computer systems and computer programs for performing one or more of the above method(s).
Abstract:
A system for pH gradient ion exchange LC-MS and methods of using such system are described. A buffer system for pH gradient LC-MS that are compatible with a mass spectrometer and methods of using such buffer system are also disclosed.
Abstract:
The invention is directed to methods for purifying compositions comprising a protein of interest, preferably for use in the clinical setting.
Abstract:
Materials, apparatuses, methods, and systems for isolating and identifying mixture components on the basis of isoelectric point ("pI"). In one aspect, the materials are particles adapted to have a predetermined isoelectric point. The particles include a porous body into which a plurality of cavities extend. A substance of a predetermined isoelectric point is deposited in the cavities to provide thereby the particle's isoelectric value.
Abstract:
This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as proteins and protein fragments from biological samples from in vivo and in vitro sources. Agents comprise a detectable group bound to a photoreactive group. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed. The invention also relates to targets isolated with these conjugates which may be useful as pharmaceutical agents or compositions that can be administered to humans and other mammals.
Abstract:
The invention provides mixed bed multi-dimensional liquid chromatographs and methods of making and using them. The invention provides systems comprising the mixed bed multi-dimensional liquid chromatographs of the invention operatively linked to mass spectrometry devices. The invention provides novel systems and methods for determining polypeptide profiles and protein expression variations, as with proteome analyses. The present invention provides systems and methods for simultaneously identifying and quantifying individual proteins in complex protein mixtures by selective differential labeling of amino acid residues followed by chromatographic and mass spectrographic analysis. The invention also provides computer program products and computer implemented methods for practicing the systems and methods of the invention.
Abstract:
The invention provides methods of monitoring the production of a target polypeptide by generating surface enhanced laser desorption/ionization mass spectral profiles of samples taken form multiple cell culture batches or of samples taken at different times from a given batch. The invention additionally provides methods for monitoring the purification of a target polypeptide from a mixture by generating surface enhanced laser desorption/ionization mass spectral profiles of samples taken at various times during a purification process. In addition, the invention relates to methods of identifying conditions that can be used in increasing the scale of a given purification process.
Abstract:
A purification device and purification method is described. The device (25) includes a quenching reagent (70) coupled to an insoluble support (60). A target compound (10) mixture flows across the quenching reagent (70), immobilizing impurity residues (20) on the quenching reagent (70) and purifying the target compound (10).