Abstract: A method for identifying a protein antigen to a target fungus is disclosed. The method comprises screening expressed proteins from a cDNA gene expression library with antisera to the target fungus and cross-screening with antisera to a nontarget fungus. Antibodies to the protein antigen are also disclosed. Methods for detecting the presence or absence of the antibodies or of the protein antigen are also disclosed, as well as kits for performing such assays. In preferred embodiments, the target fungus is H. capsulatum.

Abstract: The invention relates to monoclonal antibody capable of specific binding to the molecular weight of 20 kDa variant of human growth hormone. This monoclonal antibody has no substantial binding to hGH of molecular weight 22 kDa. The invention also relates to the use of this monoclonal antibody for measurement of hGH 20K, especially in body fluids. The antibodies can be used for detection and quantification of hGH 20K, especially in serum.

Abstract: The invention concerns the stabilization and amplification of electrochemiluminescence signals in detection methods.

Abstract: One-step enzyme immunoassays in which enzyme-antibody conjugate or label and enzyme substrate are separated until separation of bound and free enzyme conjugate or label is complete. This separation is accomplished by using variable flow paths, immobilization of substrate at the test line, placement of substrate in a sac or association with a particle label, enzyme product chemical capture, delay zone dissolution and protected enzyme substrates.

Abstract: Methods, systems, kits for carrying out a wide variety of different assays that comprise providing a first reagent mixture which comprises a first reagent having a fluorescent label. A second reagent is introduced into the first reagent mixture to produce a second reagent mixture, where the second reagent reacts with the first reagent to produce a fluorescently labeled product having a substantially different charge than the first reagent. A polyion is introduced into at least one of the first and second reagent mixtures, and the fluorescent polarization in the second reagent mixture relative to the first reagent mixture is determined, this fluorescent polarization being indicative of the rate or extent of the reaction.

Abstract: The present invention relates to the substantial elimination of errors attributable to carryover microspheres, doublets, or misclassification of microsphere subsets. The present invention is based upon passing a sufficient minimum number microspheres through the flow analyzer during an assay run.

Abstract: The hypervariable region (E2HV) of the putative hepatitis C virus (HCV) glycoprotein E2/NS1, between about amino acid 384 to about amino acid 414, is a rapidly evolving region of HCV, and is likely to be under positive immune selection. A newly discovered motif within this hypervariable region is immunogenic and conserved with respect to the character of the amino acids. In many isolates, this motif falls between amino acids 401 to 406 or 407. The discovery of this motif allows for additional materials and methods to treat and diagnose HCV.

Abstract: A method of assaying for an analyte which occurs at least partially bound as a complex with its soluble receptor or binding protein, the method comprising the steps of: i) mixing a biological fluid serum sample containing the analyte to be determined with a detergent for dissociating said complex, ii) mixing the sample from step i) with reagents, including a specific binding partner of the analyte for binding to the analyte, for performing a specific binding assay for the analyte, iii) and mixing the sample from step i) with a sequestrant for the detergent, whereby the binding of step ii) is performed in the presence of the sequestrant.

Abstract: The present invention relates to novel methods and devices for detecting whole or non-fragmented parathyroid hormone (wPTH) in a biological sample. In particular, a novel monoclonal or polyclonal antibody or antibody fragment is used that is specific for a portion of the initial peptide sequence for wPTH which comprises a domain for adenylate cyclase activation, (amino acids 2 to 8), wherein at least four amino acids in this sequence are part of the reactive portion with the antibody. The present wPTH assay can differentiate between the complete 1 to 84 amino acid sequence form of PTH and a large but incomplete 7 to 84 amino acid sequence form of PTH that is measured by commercially available “intact” or I-PTH assays. Measurement of both the complete biologically active form of PTH and the large but biologically inactive form of PTH can lead to the misdiagnosis of the parathyroid function in patients.

Abstract: Specific genetic deletions are identified in mycobacteria isolates, including variations in the M. tuberculosis genome sequence between isolates, and numerous deletion present in BCG as compared to M. tb. These deletions are used as markers to distinguish between pathogenic and avirulent strains, and as a marker for particular M. tb isolates. Deletions specific to vaccine strains of BCG are useful in determining whether a positive tuberculin skin test is indicative of actual tuberculosis infection. The deleted sequences may be re-introduced into BCG to improve the efficacy of vaccination. Alternatively, the genetic sequence that corresponds to the deletion(s) are deleted from M. bovis or M. tuberculosis to attenuate the pathogenic bacteria.

Abstract: The present invention relates to the identification of candidate ligands which modulate antigen presenting cells. Specifically, ligands which superactivate antigen presenting cells are identified.

Abstract: An increased risk of a fetal chromosomal abnormality, for example, fetal Down syndrome can be detected by separating or discriminating &agr;-fetoproteins present in the body fluid of a pregnant woman, and measuring the proportion of one or more of the &agr;-fetoproteins which have a specific sugar chain structure, relative to the total &agr;-fetoproteins.

Abstract: The present invention discloses a method for the measurement of hydride using a chemiluminescent compound. The preferred chemiluminescent molecule is an acridinium compound. The source of hydride for the reduction of acridinium compound may be of chemical or biochemical origin, or the result of enzymatic catalysis. The chemical source of hydride, for example, might be metal hydrides, such as NaBH4. A biochemical source of hydride might be that derived from NADH, or NADPH, while an enzymatic source would be the class of oxidoreductases termed dehydrogenases which convert NADH or NADPH from NAD or NADP. There are numerous potential applications for acridinium compounds as chemiluminescent indicators of hydride. Any applied tests or diagnostic assays, in which hydride is either present at the onset of or generated through the course of a reaction, would benefit from the present invention.

Abstract: The present invention provides a method by which UTI concentration can be measured easily with high precision and good reproducibility. The measurement is performed by adding free anti-UTI antibodies to a sample and measuring the degree of the resulting agglutination, for example, from the change in absorbance. As shown in FIG. 3, the UTI concentration and the degree of the agglutination (i.e. the change in absorbance) are correlated. The absorbance can be measured by using a general spectrophotometer, preferably at a wavelength of 300 to 400 nm. Polyethylene glycol is preferably added to the reaction solution as an agglutination accelerator. The polyethylene glycol preferably has an average molecular weight of 2,000 to 20,000, and the concentration of polyethylene glycol in the reaction solution is preferably in the range of 2 to 10 weight %.

Abstract: The invention provides assay methods and kits that in general measure the level of a first analyte in a sample reduced by the level of a second analyte present in the same sample. In one embodiment, where levels of a first analyte from a first source is desirably determined and first analyte in the sample released from a second source is accompanied by proportional co-release of a second analyte, the assay identifies the level of first analyte released only from the first source. For analytes within bodily fluids, the assay can differentiate between elevated levels of analyte specific to the particular physiological or pathological state and elevated levels not specific to the particular state, providing single tests with diagnostic utility.

Abstract: The present invention provides methods and compositions for the treatment of Alzheimer's disease. In particular, the present invention provides methods and compositions suitable to assess, characterize, and identify inhibitors of neuronal transport of Alzheimer's amyloid precursor protein.

Abstract: The present invention provides a method for determining viral load in a patient infected with human immunodeficiency virus, which is useful in patients where viral loads are not detectable in plasma. The levels of human immunodeficiency virus are measured in CD4−CD8− double negative cells. Furthermore, the invention also provides a kit for determining viral load in a patient infected with human immunodeficiency virus.

Abstract: Sandwich assays for collagen degradation products are conducted using an antibody of the same specificity on both sides of the sandwich or using a first antibody reactive with an epitope contained in the sequence EKAHDGGR and a second antibody which may be the same or different.

Abstract: The efficiency and accuracy of one-step lateral flow assays can be improved by employing more efficient binding between participants in labeling and capture. Thus, in addition to analyte/anti-analyte interactions, specific binding is achieved through members of an irrelevant specific binding pair. Also included within the invention is a format wherein unlabeled competitor for analyte serves as a gatekeeper in the capture zone, competing with analyte for labeled anti-analyte, which analyte will be captured in a detecting portion of a capture zone.

Abstract: The isolation, cloning and characterization of a human gene related to but distinct from the EGF receptor gene has been described. Nucleotide sequence of the gene and amino acid sequence of the polypeptide encoded by the gene have been determined. The use of the nucleic acid probes and antibodies having specific binding affinity with said polypeptide for diagnostic and therapeutic purposes has also been described.

Abstract: Disclosed is a device and method of use for detecting polyvalent analytes such as antibody to the AIDS virus, utilizing an inverse sandwich method. The test device comprises a first substance having an epitope, bound to a label and capable of moving within the test device. The test device further comprises a second substance immobilized to the test device and spatially separated from the first substance. The second substance has an epitope substantially similar to the epitope of the first substance. Upon application to the test device, the polyvalent analyte binds to the first substance and moves within the test device to the location of the second substance with both polyvalent analyte and first substance are immobilized at location of the second substance. Polyvalent analyte is detected by the presence of the label at the location of the second substance. Also disclosed is a control substance for use with the device that can be used to determine completion of the test and viability of the device.

Abstract: The invention provides multifunctional fusion constructs which are rapidly incorporated into a macromolecular structure such as a phycobilisome such that the fusion proteins are separated from one another and unable to self-associate. The invention provides methods and compositions for displaying a functional polypeptide domain on an oligomeric phycobiliprotein, including fusion proteins comprising a functional displayed domain and a functional phycobiliprotein domain incorporated in a functional oligomeric phycobiliprotein. The fusion proteins provide novel specific labeling reagents.

Abstract: The present invention relates to novel methods and devices for detecting non-complexed prostate specific antigen (PSA), which can be used either alone or in conjunction with total PSA tests to identify patients having either benign prostatic diseases (BPD), such as benign prostatic hyperplasia, prostatitis, or glandular atrophy or prostatic adenocarcinoma (CAP). In a biological sample, one can find not only non-complexed PSA, but also PSA which has formed a complex with &agr;1-antichymotrypsin (ACT). The present invention removes or precipitates complexed PSA (PSA-ACT) and ACT from a fluid sample, thereby removing any possible interference due to the binding of complexed PSA to assay reagents.

Abstract: The present invention involves a method for assaying a substance. The method of the present invention comprises contacting the substance with an assay agent comprising a catalytic agent to associate the substance with the catalytic agent, contacting the resulting associated substance with a label precursor capable of reacting catalytically with the catalytic agent to release the label, and detecting a mass label. The present invention also involves a kit for assaying a substance. The kit of the present invention comprises an assay agent comprising a catalytic agent, and a label precursor capable of reacting catalytically with the catalytic agent to release a mass label.

Abstract: A method for assaying specific binding between a fluorophore-labeled probe and an unlabeled target is provided. The method includes detecting a quenching effect on fluorescence emitted by the fluorophore-labeled probe resulting from binding. The method is conducted without separating complexes of the target and probe from the free target and free probe prior to quenching effect detecting, and without providing a signal quenching agent to quench fluorescent light. Preferably, the probe and target are amino acid-containing compounds, such as proteins. The method can be used for a variety of applications, including screening for drug candidates having optimum binding properties, and quantifying and classifying the binding characteristics between peptide-containing compounds. The method is more sensitive than conventional assays, enabling the analysis of minute samples and low affinity binding interactions between receptors and ligands that are below the detection limits of conventional technology.

Abstract: Methods for generating multiple rounds of combinatorial libraries, which use Monte Carlo methods to search the multi-dimensional composition and non-composition space of variables in combinatorial chemistry; the combinatorial libraries generated and screened by such Monte Carlo methods; and an apparatus for generating and screening such libraries robotically. The process involves preparing a first set of samples, then changing the composition and non-composition variables of the samples using Monte Carlo sampling methods, and accepting proposed new samples according to a detailed balance acceptance criterion.

Abstract: The present invention provides a method capable of simultaneous processing of plural test samples for the receptor binding property of a chemical substance, which does not require immobilization of the receptor or a special device, and a reagent to be used for this method. That is a method for assaying the receptor binding property of an assay target substance is provided, the method comprising the steps of (a) competitively reacting a known concentration of a ligand and the assay target substance with a known concentration of the receptor in a solution, (b) measuring, without physically removing the ligand bound with the receptor prior to the assay, the amount of a free ligand in the solution using one or more antibodies against the ligand, and (c) determining the receptor binding property of the assay target substance using the amount of the free ligand as an index.

Abstract: The invention concerns a method for the purification of a target substance from a biological sample by immobilizing the target substance on a solid phase by means of a high affinity binding pair and subsequently eluting it by adding a partner of the binding pair in a free form. In addition reagent kits for carrying out the method are disclosed.

Abstract: The invention relates to (1) pressure-mediated dissociation of an analyte complexed with an endogenous binding partner to enable detection of a complex formed from the analyte and an exogenous binding factor, (2) pressure-mediated association of an analyte and an exogenous binding partner to enable more rapid and/or more sensitive detection of an analyte, and (3) pressure-mediated association and dissociation of biomolecular complexes to enable separation of one biomolecule from a complex mixture. Pressure can be used to improve assays by dissociating endogenous analyte complexes and improving assay speed and sensitivity by associating the analyte molecules with exogenously supplied binding partners. Pressure can also be used to improve the separation of compounds from contaminated mixtures.

Abstract: The gene for Streptococcus pyogenes DNase B has been cloned and vectors incorporating the cloned DNA have been used to transform Escherichia coli, allowing efficient and rapid production of the DNase in E. coli without the necessity of growing large quantities of S. pyogenes. The enzyme can be produced with a leader peptide at its amino terminus. An improved method for the purification of naturally occurring S. pyogenes DNase B enzyme is also provided. The DNase B enzyme produced, either by purification of naturally occurring enzyme or by recombinant DNA techniques, can be used to generate antibodies and can also be used in immunochemical assays to detect the presence of anti-DNase B antibodies in serum as a marker of infection by S. pyogenes.

Abstract: The invention pertains to an immunoassay, and a kit for said assay, in which at least one monoclonal antibody or polyclonal antibody specifically binds to an epitope corresponding to amino acids 145 to 234 of human chromogranin A.

Abstract: The invention relates to novel methods of detecting alterations in cell cycle regulation in a cell or a cell population and screening for agents capable of modulating cell cycle regulation through the use of multiparameter assays and a fluorescence-activated cell sorter (FACS) machine.

Abstract: A magnetic focusing immunosensor for the detection of pathogens comprising a laser, an exciting fiber and a collecting fiber, a fiber optic magnetic probe in communication with the collecting and exciting fibers and means for detecting, collecting and measuring fluorescent signals in communication with the collecting fiber. The probe and the collecting and exciting fibers are configured to focus paramagnetic microspheres attached to antigen/antibody/optically labeled complexes in a predetermined pattern in the field of view of the collecting fiber while blocking background interference.

Abstract: The present invention provides a method for detecting a molecular event, comprising (1) applying an electromagnetic test signal to a sample in which a molecular event is being detected, whereby the sample interacts with and modulates the test signal to produce a modulated test signal, and (2) detecting the modulated test signal, wherein the applying and detecting take place in a temperature-controlled environment, wherein the temperature-controlled environment comprises the sample, a radiating portion of a signal generating circuit, and a receiving portion of a signal detection circuit and wherein the applying and detecting take place in the environment at a temperature controlled to within ±0.5° C.

Abstract: Methods for detecting pregnancy in a woman comprise screening a biological sample of the woman for pregnancy markers. The methods of the invention include chemiluminescent assays for the pregnancy markers. The methods of the invention also comprise utilizing at least two capture antibodies that specifically bind different epitopes of the pregnancy marker in one assay. The methods of the invention permit detection of pregnancy within about 7 days after ovulation or implantation.

Abstract: The current invention relates to new HIV-1 group O antigens, nucleic acids encoding them, and the use of said antigens and/or nucleic acids as reagents in the diagnosis and prophylaxis of AIDS. It also relates to new HIV-1 group O strains comprising these antigens.

Abstract: A monitor having a hollow semipermeable casing with a test strip is agitated in a sample volume of suspect liquid. The monitor is partially filled with tiny colored beads confined to its interior by the size of the openings in the semipermeable structure. Adhered to the beads are antibodies matched to the antigen characterizing the suspect substance. The test strip is segregated into collection and control regions. Antibodies matched to the suspect antigen are adhered only to the collection region. If the sample fluid contains the suspect substance, it will bind to both the bead antibodies and the collector antibodies, resulting in an accumulation of colored beads in the collection region but not the control region. A significant color differential between the two is a positive indication of the suspect substance. This invention has significant potential in screening for the date rape drugs, GHB and rohypnol.

Abstract: The present invention provides assays for identifying the levels of both protease sensitive and protease resistant conformers of PrPSc in a sample. In a preferred embodiment, the assay comprises determining levels of total PrPSc in a sample, subjecting the PrPSc fraction to treatment with a protease that selectively hydrolyzes the protease sensitive PrPSc (sPrPSc) conformers, and quantifying the levels of sPrPSc in the sample. The ability to detect sPrPSc allows early detection of prions, since the PrPSc in easily accessible biological samples such as blood is predominantly sPrPSc. The ratio of sPrPSc to rPrPSc also allows the identification of a particular prion strain in an infected sample.

Abstract: This invention relates to a method for establishing and using a decision marker by which positive samples can be discriminated from negative samples. The method employs the analysis of multiple samples from confirmed positive and negative samples. A fluorescence channel is selected so that the desired sensitivity and specificity are achieved. A microparticle having this fluorescence channel then is made and is used in conjunction with a fluorescence marker which is specific for the population of interest.

Abstract: The present application describes novel uses of ruthenium bipyridyls or palladium porphyrins as photo-activatable crosslinking agents. Crosslinking can be between any two molecules including peptides, proteins, or compounds. Crosslinking occurs in the presence of an electron donor such as ammonium persulfate, and requires only moderate intensity visible light. Crosslinking can be between peptides, polypeptides or lead candidate compounds to unknown target molecules. Reagents utilyzing ruthenium bipyridyls and palladium porphyrins crosslinkers for use in diagnostic and detection scenarios are also disclosed.

Abstract: A method for predicting animal growth performance, which includes administering a suitable amount of growth hormone releasing hormone (GHRH) to an animal, observing levels of growth hormone (GH) subsequent to GHRH administration and; predicting growth and fat development in an the animal from observed levels of GH.

Abstract: Antibodies which bind with antigens of human immunodeficiency virus type 1 (HIV-1), such as Lymphadenopathy Associated Virus (LAV), are disclosed. Retroviruses associated with Acquired Immune Deficiency Syndrome (AIDS) are isolated from the sera of patients afflicted with Lymphadenopathy Syndrome (LAS) or AIDS. Viral extracts, structural proteins and other fractions of the retrovirus immunologically recognize the sera of such patients.

Abstract: A method for analyzing a liquid test sample by electrochemiluminescence. The method includes at least one specific biochemical binding reaction that leads to the formation of a complex which contains a chemiluminescence marker and the binding of the complex to a magnetic microparticle. Detection is carried out in a measuring cell having a working electrode in order to determine the concentration of the marked microparticle. The detection cycle includes a purification step, a conditioning step, a recovery step and a measuring step. A specified potential profile is applied to the working electrode during these steps. Between the conditioning step and the recovery step, an additional potential pulse with an oxidizing and/or a reducing potential is inserted into the voltage shape of the detection cycle in order to improve the deposit of the microparticle.

Abstract: Methods of detecting antibodies to one or more glycosphingolipid(s) of interest in a sample are disclosed which comprise using a solid-phase reactant having carbonyl groups attached thereon, and the glycosphingolipid(s) of interest linked to the solid-phase reactant by an amide bond between an amino group of the glycosphingolipid of interest and a carbonyl group attached to the solid-phase reactant. The methods of detecting antibodies to glycosphingolipid(s) of interest can be used in methods of diagnosing autoimmune diseases in an individual.

Abstract: Unique recombinant antigens representing distinct antigenic regions of the Hepatitis C Virus (HCV) genome which can be used as reagents for the detection of antibodies and antigen in body fluids from individuals exposed to HCV. The present invention also provides an assay for detecting the presence of an antibody to an HCV antigen in a sample by contacting the sample with the recombinant antigens. Preferred assay formats include a screening assay, a confirmatory assay, a competition or neutralization assay and an immunodot assay.

Abstract: A method for determining the affinity between binding partners, or a property of one of the binding partners dependent on the affinity, comprising the steps of: (i) contacting the binding partners, one of which is immobilised on a surface; (ii) oscillating the surface at increasing amplitude; and (iii) detecting a dissociation event. An analogous method can be used to separate a target analyte from a composition. The subject invention also pertains to an apparatus for determining the affinity between binding partners, and comprises: a surface (10) having one binding partner (16) immobilised thereon; means for oscillating the surface at increasing amplitude; and a device (14, 15) for detecting a dissociation event.

Abstract: The present invention relates to methods for identifying nucleic acid molecules encoding (poly)peptides that interact with target molecules. The method of the present invention is particularly characterized by an in vitro translation step under conditions that allow formation of polysomes in the presence of antisense oligonucleotides complementary to the tag-coding sequence of ssrA-RNA. The present invention further relates to kits that are useful for carrying out the method of the invention.

Abstract: The invention provides improved immunoassay techniques for detecting the presence of analytes in a liquid sample. The present immunoassay methods utilize anti-allotypic monoclonal antibodies as capture reagents for primary binding proteins specific for the analytes of interest. The monoclonal antibodies are highly specific for the allotypic determinants present on the primary binding protein. The use of anti-allotypic monoclonal antibodies as capture reagents provides improved levels of specificity and accuracy of the immunoassay, in part because interference from endogenous immunoglobulins in the sample is significantly reduced. The invention further provides anti-allotypic monoclonal antibodies.

Abstract: A method for measuring the activity of creatine kinase MB (CK-MB) isozyme which is accurate, high in specificity, convenient, by inhibiting the activity of a mitochondria localized creatine kinase (mCK) isozyme to avoid the influence of mCK and a measurement reagent therefor.

Abstract: A mass biosensor uses an intermediate avidin layer to facilitate binding of a biotinylated antibody to a measurement surface of the biosensor. The avidin layer can be added by the manufacturer of the biosensor, while the biotinylated layer can be added by the user. This two-phase method of chemically modifying the measurement surface significantly reduces the user time required to customize the measurement surface to render it capable of binding selected compounds. An organosilane coupling agent attached to the surface provides sites to which avidin is bound. Avidin acts as a universal receptor of biotinylated compounds with specific binding affinities. Biotinylated antibodies or other biotinylated compounds are added and bind to the immobilized avidin. Surface adsorption is reduced by washing the modified surface with biotin to block potential sites of weak bond formation, electrostatic and hydrophobic interactions.