Abstract: A family of GFP scaffolds capable of accommodating two proximal, randomized binding loops is disclosed. GFP-based binders binding with nanomolar affinity are developed from a library of these GFP scaffolds.

Abstract: The invention provides a novel class of cyanine dyes that are functionalized with a linker moiety that facilitates their conjugation to other species. Also provided are conjugates of the dyes, methods of using the dyes and their conjugates and kits including the dyes and their conjugates.

Abstract: The invention provides a novel class of fluorescent compounds. Also provided are conjugates of the fluorescent compounds, methods of using the fluorescent compounds and their conjugates as well as kits including the fluorescent compounds and their conjugates.

Abstract: The invention provides novel Class I HLA-A2 and Class II HLA-DR4-restricted epitopes and methods for their use in detecting T-cells in peripheral blood specific for infection or latency of mycobacterial infection, including M. tuberculosis and M. leprae as others. For example, methods for diagnosing the presence of infection or exposure by M. tuberculosis utilize multimers of HLA monomers or modified monomers having a bound HLA-binding peptide to perform high throughput screening of patient PBLs. The methods can be used for monitoring the success of anti mycobacterial treatment in patients and to screen vaccines and drugs for effectiveness in treating or preventing exposure, infection and latency of mycobacteria in humans.

Abstract: Disclosed herein are immunogenic compositions comprising a multilayer film comprising two or more layers of polyelectrolytes, wherein adjacent layers comprise oppositely charged polyelectrolytes. A first layer polyelectrolyte comprises an antigenic polypeptide comprising one or more surface adsorption regions covalently linked to one or more antigenic determinant regions, wherein the antigenic polypeptide and the one or more surface adsorption regions have the same polarity. The immunogenic compositions may be employed in methods of eliciting an immune response in a vertebrate organism.

Abstract: Disclosed are sequences encoding monomeric variants of DsRed fluorescent proteins and methods of use.

Abstract: This invention provides dye phosphoramidites, particularly phosphoramidites of substituted cyclic bridged cyanine and related dyes, of the general formula: In this formula, each dotted line represents carbon atoms necessary to form a fused substituted or unsubstituted aromatic ring; m is an integer from 1 to 18; Y and Z are independently selected from the group consisting of S, O, N, CH2 and C(CH3)2; R1 is an alkyl; (PAM) is a phosphoramidite group; X{circle around (?)} is a negative ion; and Q is L-W, wherein L is a conjugated cyclic moiety and W is OR2, wherein R2 is a second alkyl. Methods of making and using the dye phosphoramidites are also provided.

Abstract: The invention provides a method for detection of receptor- or ion channel-modulators (e.g., antagonists, blockers, etc.). The method comprises fractionating a sample by using a liquid-based separation means (e.g., such as a capillary electrophoresis device), and feeding the fractions containing the modulators directly to a biosensor. The biosensor is activatable by a receptor agonist and produces a measurable response as a result of this activation. Preferably, the agonist is fed to the biosensor through the liquid-based separation means, together with fractions containing the modulators (e.g., antagonists or blockers). The presence of a modulator is detected by monitoring a change in the response of the biosensor, e.g., by measuring changes in electrical properties of the biosensor (such as through patch clamp analysis).

Abstract: In a method of determining heparin content, a known amount of thrombin (FIIa) or activated coagulation factor X (FXa) is added to a mixture which comprises a known concentration of chromogenic substrate (S), a known concentration of antithrombin (AT) and a sample having an unknown heparin concentration. The amount of FIIa or FXa is chosen such that at most 20%, of the S present is reacted during the period which the AT needs to deactivate all the FIIa or FXa present. The final concentration of the reacted chromogenic substrate p-nitroanilide ([pNA].sub.final) is determined after completion of the reactions, and the [pNA].sub.final is used to determine the rate constant (k.sub.dec) of the reaction of FIIa or FXa with AT using the relationship: ##EQU1## The heparin concentration in the sample can then be determined using the value of k.sub.dec from a predetermined calibration curve of k.sub.dec against heparin concentration.

Abstract: It is intended to present a sensor capable of labeling with more dyes, applying the labeled protein in immunochromatography making use of antigen-antibody reaction, and having an excellent sensitivity. In a buffer, a protein and a first covalent bonding compound that can react with this protein are reacted to prepare a protein conjugate, then a cyanine labeling dye is added in the buffer containing the protein conjugate, and the protein conjugate and cyanine labeling dye are reacted to prepare a dye labeled protein conjugate. Alternatively, in a buffer solution, a protein and a cyanine labeling dye are reacted to prepare a dye labeled protein, then a first covalent bonding compound that can react with the protein is added in the buffer containing the dye labeled protein, and the dye labeled protein and first covalent bonding protein are reacted to prepare a dye labeled protein conjugate.

Abstract: Water soluble glycoconjugated fluorescent labeling reagents comprised of a fluorescent dye, one or more sugar or carbohydrate residues that impart water solubility, and a reactive group that allows covalent attachment of the reagent to a substrate are disclosed. Glycoconjugated fluorescent labeling reagents are depicted as follows: ##STR1## These glycoconjugates are highly water soluble because of the sugar residues. This property is expected to lower nonspecific binding to cellular matter, reduce precipitation of labeled substrates, inhibit quenching of fluorescence, and unlike many prior fluorescent labeling reagents, that are soluble only in organic solvents, render the glycoconjugates usable in wholly aqueous solution.

Abstract: The synthesis of 10,10'-substituted-9,9'-biacridine molecules and their derivatives is disclosed. These molecules are shown to catalyze the production of light by chemiluminescence in the presence of a signal solution having at a pH from about 10.0 to about 14.0, at a concentration effective for producing a chemiluminescent signal, a chelating agent, a sulfoxide, a reducing sugar, an oxidant or combination of oxidants, an alcohol and aqueous sodium tetraborate. These 10,10'-substituted-9,9'-biacridines are used alone or attached to haptens or macromolecules and are utilized as labels in the preparation of chemiluminescent, homogeneous or heterogeneous assays. They are also used in conjunction with other chemiluminescent label molecules to produce multiple analyte chemiluminescent assays.

Abstract: The present invention provides a method of increasing the duration of detectable photon emission of a luciferase-luciferin reaction. The method provides a luciferase-luciferin reaction in which photon emission can be detected for up to and including eight hours. A method of the present invention can also be used to detect the presence of luciferase in biological samples. The present, invention also provides a composition used in detecting the presence of luciferase in biological samples.

Abstract: A fusion protein suitable for use as a vaccine comprises an amino acid sequence having biological activity which is fused via an intervening hinge comprising from two to eight glycine-proline repeats to the C-terminus of sufficient of the amino acid sequence of a B subunit of an enterotoxin which is capable of ADP-ribosylation of a GTPase.

Abstract: Photochromic compositions comprise a bacteriorhodopsin suspension, at least one organic nitrogen-containing compound and a binder. The composition may further include a detergent. Photochromic materials comprise a support and a photochromic film formed on the support from a photochromic composition as described.

Abstract: This disclosure related to a method and reagents for determining tetrahydrocannabinoids (THC) and THC metabolites in a biological fluid such as urine. In particular, this disclosure relates to a fluorescence polarization immunoassay procedure for determining the presence of THC and to a novel class of tracer compounds employed as reagents in such procedures. The procedure described also provides for novel wash reagent for a THC fluorescence polarization assay.

Abstract: The present invention relates to iminium ion substituted cyanine dyes having a fluoresence absorbance of between about 500 and 900 nm, a reduced tendency to aggregate and enhanced photostability. The cyanine dyes of the present invention are represented by the formula ##STR1## wherein n is 0, 1, 2 or 3;m is 0, 1, 2 or 3;R.sub.1 and R.sub.2 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;R.sub.3 and R.sub.4 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;R.sub.5 and R.sub.6 are independently selected from the group consisting of (CH.sub.2).sub.p X where p is 1-18 and X is a functional group that reacts with amino, hydroxyl and sulfhydryl nucleophiles;R.sub.7 and R.sub.8 are independently selected from the group consisting of hydrogen, C1-C10 alkyl groups and where R.sub.7 and R.sub.8 are taken together to form a five- or six- membered heterocyclic ring;R.sub.

Abstract: Disclosed is an assay system including a compound comprising an analyte-specific moiety having substituted thereon a polymer comprising plurality of self-quenching emitter moieties and a plurality of isocharged functionality separating the emitter moieties. The present invention provides compounds that overcome the undesirable effects of self-quenching when multiple emitter moieties are used for labelling of assay reagents. Avoidance of this self-quenching phenomenon by the compounds of the invention makes it possible to introduce a more concentrated degree of labelling on to analyte-specific molecules such as oligo nucleotide probes, antibodies and other specific binding proteins and analyte-specific polysaccharides. Therefor, it is possible to effect greater assay sensitivity because the number of labels per recognition molecule(analyte-specific moiety) can be increased beyond the point previously possible without the reduction in signal caused by self-quenching.

Abstract: The subject invention provides a method for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The method for analyzing metabolic activity of cells involves the use of a substrate having the formXR-REPORTER-BLOCKwherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate,-REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properities different from those of the substrate, andXR-- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z--S--H) to form a thioether conjugate (Z--S--R--).

Abstract: A method for the generation of phenylthiocarbamyl (PTC) amino acids from phenylthiohydantoin (PTH) or anilinothiozolinone (ATZ) amino acids. The method involves a base-catalyzed ring opening of the PTH or ATZ in the presence of a reducing agent. The method affords an alternative to the established aqueous acid conversion reaction of the Edman degradation in which ATZ and PTC amino acids are converted to the PTH amino acid. A further method is described for the generation of reactive ATZ amino acids from PTH amino acids. These methods facilitate the analysis of protein at low molar amounts by allowing the synthesis of amino acid derivatives which can be analyzed in quantities which are much lower than those required for conventional PTH amino acid analysis.

Abstract: A composition for testing periodontal diseases which diagnoses or prognosticates contraction or progress of the diseases or diagnoses the therapeutic value by promptly determining peptidase-like enzymatic activity in a specimen. The composition is a combination of a compound of the formula [1] or [2] or a mixture thereof, a chromogen and an oxidase:X-T-Pro-Y [1]orX'-Z'-Arg-Y' [2]wherein Pro is proline residue; Arg is arginine residue; X and X' are hydrogen or an amino protecting group, respectively; Y and Y' are a residue of a compound which can increase oxidation reaction rate of a chromogen with a oxidase in the presence of oxygen and is attached to the C-terminal of Pro or Arg, respectively; and T and Z' are an amino acid or peptide residue composed of 0 to 4 amino acids or their protected derivatives the C-terminal of which is attached to the N-terminal of Pro or Arg, respectively.

Abstract: Activities of physiologically active substances such as endopeptidases, proteases, transferases, phosphatases, protein kinase and the like enzymes can be measured specifically with high sensitivity by using as a substrate a peptide derivative having 2 to 15 amino acid units and a group which can emit fluoresence and/or absorb ultraviolet light as at least one terminal group.

Abstract: A method for obtaining DNA expressing histidine rich protein of various types of Plasmodia is disclosed. The method involves hybridization with the comparable DNA of P. lophurae. The method of particularly well suited for obtaining P. falciparum DNA, whether it is associated with know or knobless phenotype. Additionally, the invention disclosed a safe method for diagnosing P. falciparum infection.

Abstract: The invention relates to a process for the preparation of purple membrane containing bacteriorhodopsin, which process comprises obtaining, in a manner know per se, the cell membrane from halobacteria cells, and subjecting the material to gel filtration chromatography in order to isolate the purple membrane from the cell membrane.

Abstract: Chromogenic substrates for the detection of hydrolyzing enzymes, processes for the preparation of these chromogenic substrates and the use of the chromogenic substrates.The quantification of hydrolyzing enzymes for diagnostic purposes has to date been carried out by determination of the amounts of a fluorescent or highly absorbent chromogenic dyestuff liberated in the hydrolytic reaction, detection of which has in part been possible exlusively by instruments or in the neutral to alkaline pH range. After enzymatic hydrolysis, the chromogenic substrates according to the invention lead to highly sensitive and specifically measurable signals regardless of the pH range.Azo dyestuff compounds with the general formulaA--N.dbd.N--B (OR)in which A) and B) have the meanings given in claim 1 andR) is a radical which can be liberated by enzymatic hydrolysis, excluding a carbonyl radical,are provided.After enzymatic hydrolysis, highly sensitively and specifically measurable signals are obtained regardless of the pH range.

Abstract: A synthetic peptide of the formula: H-.sub.D -A.sub.1 -Leu-Lys-NH ##STR1## wherein A.sub.1 is Pro or Ala, is excellent in solubility in water and substrate specificity and is suitable as a substrate for determining trypsin and .alpha..sub.1 -antitrypsin.

Abstract: The present invention provides a dry reagent for blood coagulation tests in which an at least partial course of the coagulation cascade takes place, comprising a carrier material which contains a chromophoric substrate of a protease of the blood coagulation system, at least one factor and/or co-factor of the blood coagulation system and a buffer.

Abstract: Substantially pure cromolyn binding protein is prepared by means of affinity chromatography of cromolyn derivatives bound to insoluble matrices. Aminocromolyn is prepared by a six-step synthesis and amine derivatives thereof are prepared by conventional means. Obtaining a compound having an amine group instead of the OH group at the 2-carbon of the propane link of cromolyn permits many kinds of reactions without interfering with the portion of the cromolyn molecule with causes its pharmacological activity. The cromolyn derivatives can be conjugated to proteins such as BSA by means of glutaraldehyde cross-linking and such conjugates can be covalently bound to agarose beads. Cromolyn binding protein can be isolated by passing lysates of RBL-2H3 cells through chromatographic columns packed with such beads. The cromolyn binding protein can be further purified by means of lectin-agarose columns.

Abstract: The present invention provides resorufin derivatives of the general formulae: ##STR1## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5, which can be the same or different, are hydrogen, halogen, carboxyl, carboxamido, lower alkoxycarbonyl, cyano or nitro groups or lower alkyl or lower alkoxy radicals, which can be substituted by carboxyl, carboxamido, lower alkoxycarbonyl, cyano or nitro groups, and wherein R.sup.4 and R.sup.5 can together also represent an anellated aromatic residue, Z is a bridge member, A is the residue of a ligand and n is a whole number of from 1 to 200.The present invention also provides processes for the preparation of these resorufin derivatives, as well as intermediates for the preparation thereof.

Abstract: A method for preparing selected peptide substrates for detecting the activity of virus-specified proteases is provided. Specific tetrapeptide substrates are disclosed which are conjugates of protease-cleavable indicator groups and peptide sequences resembling picornavirus protease cleavage recognition sites.

Abstract: A compound for use as a chemiluminescent label in immunoassay comprises an aryl acridinium ester linked to an N-succinimidyl moiety. The compound is conveniently linked to a monoclonal antibody or other protein and is used in a two-site immunoassay for the quantitation of an antigen of interest, by initiation of the luminescent reaction and subsequent measurement of the photonic emission of the immune complex formed during the immunological reaction.

Abstract: A method for enhancing the chemiluminescent signal of an acridinium ester in a chemiluminescent reaction which comprises oxidizing the acridinium ester in the presence of an enhancer selected from the group consisting of: (a) a cationic surfactant; (b) a nonionic surfactant; and (c) a sulfated primary alcohol.

Abstract: A chromophoric peptide is prepared of the formulaDIS--D--Ala-Pro-Rand salts thereof in which R represents Gly-OH, NH.sub.2 or Gly-R.sup.2 wherein R.sup.2 denotes a carboxyl protective group, and DIS denotes 5-dimethylaminonaphthalene-1-sulfonyl. The enzyme, peptidylglycine-.alpha.-amidizing monooxygenase, is quantitatively detected by contacting a sample containing the enzyme with the peptide of the above formula when R denotes Gly--OH, separating a resultant peptide of the above formula when R denotes NH.sub.2 and determining the amount thereof quantitatively. When R is Gly--R.sup.2, the protective group R.sup.2 is removed to produce the peptide when R is Gly--OH. Advantages of the peptide are that C-terminal degradation caused by carboxypeptidases is rendered more difficult by the incorporation of proline, and the peptide contains a fluorescent group.

Abstract: A method for conducting assays for an analyte, usually in a biological sample, utilizes reagents which contain biological ligands linked to fluorescein or a derivative of fluorescein through a linkage to the fluorescein nucleus selected from acetamido and thioacetamido. In these reagents, the fluorescence efficiency is improved over that obtained in the commonly used FITC-labeled reagents.

Abstract: Sensitive proteins and other macromolecules, such as enzymes, antibodies, antigens, serum complement, fluorescent proteins, vaccine components, polysaccharides such as agarose etc, can be preserved by drying at ambient temperature and at atmospheric pressure in the presence of trehalose. A porous matrix impregnated with trehalose is provided as a receiver for a blood or other liquid sample to be dried.

Abstract: Peridinin-chlorophyll-protein complexes are provided for use as fluorescent labels and are particularly useful in diagnostic assays employing as a reagent a fluorescent compound conjugated to a member of a specific binding pair, wherein the pair consists of a biochemical ligand and a receptor and the diagnostic assay comprises a step in which the conjugate binds to its complementary binding-pair member.

Abstract: Novel adducts of oxazine urea chromophors or thiazine urea chromophors with organic substrates are provided which are useful in analytical techniques for the detection and measurement of biological and clinical compounds of interest.

Abstract: Labellable reagents for fluormetric assays comprise a cyclic condensation product of a .beta.-diketone, an aldehyde and an NH.sub.2 -bearing macromolecule, for example an antigen or antibody or a substance having an active group to which an antibody or antigen is linked. The reagents can chelate lanthanide metal ions such as Eu(II) and Tb(III) to form fluorescing complexes which can be used as labelled reagents for fluorometric assay of organic substances, for example antigens, antibodies and other substances occurring in body fluids.

Abstract: Novel compounds represented by the following general formula (1) and salts thereof: ##STR1## wherein n represents an integer of 3 to 4, R.sup.1 represents ##STR2## or --SO.sub.2 R.sup.2, and R.sup.2 represents optionally branched lower alkyl group having 1 to 6 carbon atoms, phenyl group, benzyl group or tolyl group, which are useful as substrate for use in the measurement of biological components; a substrate for use in the measurement of biological component which comprises said novel compound or salt thereof; and a method for measuring biological component which comprises using said substrate.

Abstract: Tripeptide derivatives, characterized by the general formula:R.sub.1 --X--D--Arg--A--Arg--NH--R.sub.2wherein R.sub.1 is hydrogen, .alpha.- or .beta.-naphtyl residue, lower alkyl residue which may be substituted with a carboxyl group, unsubstituted or substituted phenyl- or phenylalkyl residue. ##STR1## or a single bond with the proviso that when R.sub.1 is hydrogen then and only then X is a single bondA=Gly or SarR.sub.2 =an aromatic or heterocyclic residue which gives a compound R.sub.2 --NH.sub.2 by enzymatic hydrolysis, which can be determined quantitativelyor disalts and trisalts of inorganic or organic acids thereof, process for their preparation and method for determination of serine proteases, especially Factor X.sub.a,or components which can interact with serine proteases or zymogen forms thereof.

Abstract: Peptide sequences consisting of 2-4 amino acids with high affinity and comparatively high specificity to a number of various, physiologically important proteases are known to have been synthetized before. Such sequences with an added C-terminal marker have been widely used as substrates for the quantitative determination of the kind of proteases mentioned above. The method is based on the fact that the marker is split off under influence of the enzyme and that the liberated marker possesses an easily measurable, for instance, optic property which differs from that of the original substrate. The type of markers used until today have mainly been chromophores or fluorophores which can be quantified by photometry or fluorometry.The present invention relates to a new type of markers coupled to known peptide sequences. These markers are luminol or isoluminol which in their free state can be brought to luminate intensely, but lose considerably in luminescence when they are amide-bound to a peptide sequence.

Abstract: The present invention relates to novel acridinium esters which are useful as luminescent labels in specific binding assays such as immunoassays or nucleic acid hybridization assays. More particularly, polysubstituted aryl acridinium esters are highly stable labels for use in a chemiluminescent immunoassay.

Abstract: Water soluble xanthylium derivative substrates of rhodamine 110 and rhodol permit spectrophotometric and fluorescent measurements of trypsin-like enzymes without the addition of organic solvent additives and/or special water solubilizing agents. These novel substrates exhibit increased sensitivity for determining low levels of activity of trypsin-like enzymes such as proteolytic enzymes, cofactors, activators, antiactivators, and inhibitors. These substrates can be substituted for fibrinogen or monitor the pathways of blood coagulation.

Abstract: Reagent for immunoassay, ligand binding assay and ligand receptor assay in which luciferin is covalently bonded to a molecule having biological activity for bonding to a particular biologically active group of a material and method in which the luciferin and biologically active molecule conjugate is added to a material having a group for combination with the biological activity of the conjugated molecule, the mixture is incubated to bond the material to the reagent through the biological activity and the product of the bonding is reacted with luciferase to produce bioluminescence having intensity dependent on the concentration of the material being assayed.

Abstract: A novel chromogenic and fluorescent substrate for measuring the concentration of kallikrein in urine. The novel substrate according to the present invention has very excellent substrate specificity to kallikreins and good solubility in water or biological test solutions.The present substrate is therefore useful for measuring the concentration of kallikrein in urine for the diagnosis of hypertension and other diseases caused by the lowering of the concentration of kallikrein.

Abstract: Peptide derivatives of formula ##STR1## wherein R.sup.1 represents a substituted amino group which is capable of being split off enzymatically with formation of a colored or fluorescent compound R.sup.1 -H. The said peptide derivatives are used for quantitatively assaying the enzyme C.sub.1 -esterase. The assaying is carried out by reacting a medium which contains C.sub.1 -esterase or in which the latter is formed or consumed with a peptide derivative as defined above and measuring photometrically, spectrophotometrically, fluorescence-spectrophotometrically or electrochemically the quantity of split product R.sup.1 -H released per time unit by the catalytic hydrolytic action of the said enzyme on the peptide derivative.

Abstract: A novel chromogenic and fluorescent substrate for determining the activity of blood coagulation factor Xa and Xa-like enzymes. The novel substrate according to the present invention has very excellent selectivity as compared with the hitherto disclosed substrates and enables specific determination of blood coagulation factor Xa. The present substrate is therefore useful for the studies of chemical reactions involving formation, inhibition and consumption of factor Xa and can be also used for the determination of various factors associated therewith such as factor X, anti-Xa factor, factor VII, factor VIII, factor IX and heparin.