Abstract: The invention is directed to a method of determining whether a non-purified sample contains a target protein bound to a ligand of interest comprising the steps of: a) exposing said non-purified sample to a temperature which is capable of causing or enhancing precipitation of the unbound target protein to a greater extent than it is capable of causing or enhancing precipitation of the target protein bound to said ligand; b) processing the product of step a) in order to separate soluble from insoluble protein; and c) analyzing either or both the soluble and insoluble protein fractions of step b) for the presence of target protein, wherein said target protein is not detected on the basis of enzymatic activity of a tag, peptide, polypeptide or protein fused thereto. Particularly, the invention may be used to determine whether drugs can bind to their protein targets in samples derived from patients to ascertain whether a certain drug can be used in a therapy for that patient.

Abstract: The present invention concerns a method of determining whether a non-purified sample contains a target protein bound to a ligand of interest comprising the steps of a) exposing the non-purified sample to a temperature which is capable of causing or enhancing precipitation of the unbound target protein to a greater extent than it is capable of causing or enhancing precipitation of the target protein bound to the ligand; and b) analyzing said sample for the presence of soluble or native target protein using two or more affinity reagents capable of binding to said soluble or native target protein with a higher affinity than to an unfolded and/or insoluble form of said target protein. The invention particularly concerns the use of two affinity reagents (e.g. antibodies) which are capable of distinguishing between soluble or native, and unfolded and/or insoluble forms of a target protein and whose detection e.g. by FRET based technology, allows the performance of the method without a separation step.

Abstract: The invention is directed to a method of determining whether a non-purified sample contains a target protein bound to a ligand of interest comprising the steps of: a) exposing said non-purified sample to a temperature which is capable of causing or enhancing precipitation of the unbound target protein to a greater extent than it is capable of causing or enhancing precipitation of the target protein bound to said ligand; b) processing the product of step a) in order to separate soluble from insoluble protein; and c) analysing either or both the soluble and insoluble protein fractions of step b) for the presence of target protein, wherein said target protein is not detected on the basis of enzymatic activity of a tag, peptide, polypeptide or protein fused thereto. Particularly, the invention may be used to determine whether drugs can bind to their protein targets in samples derived from patients to ascertain whether a certain drug can be used in a therapy for that patient.

Abstract: The present invention concerns a method of determining whether a non-purified sample contains a target protein bound to a ligand of interest comprising the steps of a) exposing the non-purified sample to a temperature which is capable of causing or enhancing precipitation of the unbound target protein to a greater extent than it is capable of causing or enhancing precipitation of the target protein bound to the ligand; and b) analysing said sample for the presence of soluble or native target protein using two or more affinity reagents capable of binding to said soluble or native target protein with a higher affinity than to an unfolded and/or insoluble form of said target protein. The invention particularly concerns the use of two affinity reagents (e.g. antibodies) which are capable of distinguishing between soluble or native, and unfolded and/or insoluble forms of a target protein and whose detection e.g. by FRET based technology, allows the performance of the method without a separation step.

Abstract: The invention is directed to a method of determining whether a non-purified sample contains a target protein bound to a ligand of interest comprising the steps of: a) exposing said non-purified sample to a temperature which is capable of causing or enhancing precipitation of the unbound target protein to a greater extent than it is capable of causing or enhancing precipitation of the target protein bound to said ligand; b) processing the product of step a) in order to separate soluble from insoluble protein; and c) analysing either or both the soluble and insoluble protein fractions of step b) for the presence of target protein, wherein said target protein is not detected on the basis of enzymatic activity of a tag, peptide, polypeptide or protein fused thereto. Particularly, the invention may be used to determine whether drugs can bind to their protein targets in samples derived from patients to ascertain whether a certain drug can be used in a therapy for that patient.

Abstract: The present invention concerns a method of identifying a cell colony which expresses a soluble variant of a target protein which method comprises (a) subjecting the cell colony to conditions which are capable of causing lysis thereof; (b) filtering the lysate through a filter having pores which allow only soluble proteins to pass through the filter; and (c) detecting target protein which has passed through the filter where the target is not detected an the basis of its own enzymatic activity. The invention further covers the identification of a cell colony expressing a soluble variant of a membrane protein using steps (a) and (b) of the method above. A kit for use in the methods is also covered by the invention.

Abstract: The present invention relates to an isolated leukotriene A4 (LTA4) hydrolase, which LTA4 hydrolase is present in its naturally occurring three dimensional form. It is the first three-dimensional structure of any protein component of the leukotriene cascade and enables a description of the structural basis and molecular mechanisms for the two catalytic activities of LTA4 hydrolase. Further, the invention also relates to LTA4 hydrolase complexed with an inhibitor. The structural information provided by the present invention will make possible rational design of enzyme inhibitors, which may be developed into clinically useful anti-inflammatory drugs.