This invention provides methods and compositions, e.g., to reduce interference from non-specific binding sample constituents in a migration shift assay. Interference due to non-specific binding of sample constituents to an affinity substance (e.g., an affinity molecule or a conjugate of an affinity molecule and a charged carrier molecule) is prevented by, e.g., binding the constituents to charged polymers such as heparin sulfate. The present invention also provides methods to concentrate an analyte of interest with high concentration and to detect the analyte with high sensitivity, and further to optimize the reaction conditions for easily concentrating the analyte. Such objects of the present invention are attained, for example, by concentrating a complex of the analyte and a conjugate which is formed by contacting the analyte in a sample with an affinity molecule bound to a charged carrier molecule such as DNA.

This invention provides methods and compositions, e.g., to reduce interference from non-specific binding sample constituents in a migration shift assay. Interference due to non-specific binding of sample constituents to an affinity substance (e.g., an affinity molecule or a conjugate of an affinity molecule and a charged carrier molecule) is prevented by, e.g., binding the constituents to charged polymers such as heparin sulfate. The present invention also provides methods to concentrate an analyte of interest with high concentration and to detect the analyte with high sensitivity, and further to optimize the reaction conditions for easily concentrating the analyte. Such objects of the present invention are attained, for example, by concentrating a complex of the analyte and a conjugate which is formed by contacting the analyte in a sample with an affinity molecule bound to a charged carrier molecule such as DNA.

A method of separating free target analyte from protein-bound target analyte is described. Such can include obtaining an aqueous sample containing a target analyte in a free form (free target analyte) and the target analyte in a protein-bound form (protein-bound target analyte), passing the aqueous sample through a size exclusion chromatography matrix with a molecular weight cut off sufficient to allow the free target analyte to permeate into pores of the size exclusion chromatography matrix and exclude the protein-bound target analyte, whereupon the free target analyte adheres to and is immobilized by the size exclusion chromatography matrix and the protein-bound target analyte does not adhere to by the size exclusion chromatography matrix, separating the free target analyte from the protein-bound analyte by removing the protein-bound target analyte from the size exclusion chromatography matrix, and eluting the free target analyte from the size exclusion chromatography matrix with an organic solvent.

A method of separating free target analyte from protein-bound target analyte is described. Such can include obtaining an aqueous sample containing a target analyte in a free form (free target analyte) and the target analyte in a protein-bound form (protein-bound target analyte), passing the aqueous sample through a size exclusion chromatography matrix with a molecular weight cut off sufficient to allow the free target analyte to permeate into pores of the size exclusion chromatography matrix and exclude the protein-bound target analyte, whereupon the free target analyte adheres to and is immobilized by the size exclusion chromatography matrix and the protein-bound target analyte does not adhere to by the size exclusion chromatography matrix, separating the free target analyte from the protein-bound analyte by removing the protein-bound target analyte from the size exclusion chromatography matrix, and eluting the free target analyte from the size exclusion chromatography matrix with an organic solvent.

Disclosed are a one-pot biosensor and an immunoassay method using the same. The one-pot biosensor includes a photocatalyst substrate deposited with metal nanoparticles; and a reaction pad which is disposed on an upper surface of the photocatalyst substrate and includes a first binding material-fluorescent material complex specifically binding to a molecule to be detected, and the immunoassay method using the same. The one-pot biosensor may detect a target by once solution injection and has a size enough to be portable. Accordingly, since the one-pot biosensor can detect the target by only once solution injection without a washing step, because of a sensor platform capable of being easily used by an individual other than a diagnostic expert, it is predicted to be positioned as a means capable of confirming the health condition of the individual without seeing the doctor, such as a pregnancy diagnostic kit which has been currently commercialized.

Disclosed are a one-pot biosensor and an immunoassay method using the same. The one-pot biosensor includes a photocatalyst substrate deposited with metal nanoparticles; and a reaction pad which is disposed on an upper surface of the photocatalyst substrate and includes a first binding material-fluorescent material complex specifically binding to a molecule to be detected, and the immunoassay method using the same. The one-pot biosensor may detect a target by once solution injection and has a size enough to be portable. Accordingly, since the one-pot biosensor can detect the target by only once solution injection without a washing step, because of a sensor platform capable of being easily used by an individual other than a diagnostic expert, it is predicted to be positioned as a means capable of confirming the health condition of the individual without seeing the doctor, such as a pregnancy diagnostic kit which has been currently commercialized.

Disclosed herein are compositions and uses thereof for detection of disease-related antibodies. The methods include contacting a biological sample with a solid support comprising one or more antigens that bind one or more therapeutic monoclonal antibodies, and detecting the disease-related antibody in the biological sample using an electrophoretic method.

A method for determining an amount of functional albumin includes providing a test sample containing a defined amount of albumin of unknown binding capacity and a reference sample containing the same defined amount of albumin having a reference binding capacity, incubating the test and reference samples with a defined amount of at least one albumin-binding marker M under conditions that allow formation of complexes of the at least one albumin-binding marker M and albumin (M:A), removing the complexes, detecting a presence or an amount of unbound marker M in the samples after removal of the complex (M:A) through a first and a second test strips that allow for a determination of an amount of unbound marker M, and determining the amount of functional albumin based on the presence or the amount detected of marker M.

An immunoadsorbent and a composite affinity column for purifying fumonisin B1, anguidin, T-2 toxin, zearalenone, and vomitoxin. The immunoadsorbent includes a solid phase carrier, and a fumonisin B1 monoclonal antibody, an anguidin monoclonal antibody, a T-2 toxin monoclonal antibody, a zearalenone monoclonal antibody and a vomitoxin monoclonal antibody which are coupled to the solid phase carrier, the anguidin monoclonal antibody is a monoclonal antibody secreted by a hybridoma cell strain DAS5G11E7 having an accession number of CCTCCNO:C201881. The affinity column can be used for high performance liquid chromatography-mass spectrometry detection of the fumonisin B1, the anguidin, the T-2 toxin, the zearalenone and the vomitoxin, and has stable performance. Furthermore, an economical, quick, precise and safe detection method is established of the basis of the affinity column, and can be used for purifying and detecting samples of the five toxins without mutual interference and influence.

Provided herein are compositions and methods for detecting the binding of a peptide to an MHC molecule, and the binding of a peptide:MHC complex to a TCR. In preferred embodiments, the compositions and methods are in a highly-multiplexed way. The compositions and methods disclosed herein can be used to provide direct information on which peptides are bound to an MHC molecule. Also provided is a method for simultaneously detecting a large number of peptides for binding to an MHC molecule and/or a T cell. A method for detecting competitive binding of a large number of peptides to an MHC molecule and/or a T cell is also disclosed. Also provided herein is a method for simultaneously detecting a large number of specific TCRs. The compositions and methods of the present invention are useful for vaccine design, research and monitoring of autoimmune and infectious disease, immunogenicity testing of therapeutics, and tissue typing.