Abstract: The present invention is directed to extracorporeal cell systems infected with hepatitis C virus (HCV). The present invention also relates to products of such cell systems and their use as vaccines and in immunoassays. Methods whereby HCV-infected extracorporeal cell systems are constructed are included, and various immunoassays to detect HCV antibodies are also presented. The HCV-infected cell systems can be used to screen putative antiviral agents.

Abstract: A method for generating multiple mimics of an active site of a molecule, such as a protein, using computer modeling of the active site. A putative active site is identified, and the amino acids, or residues, purportedly responsible for the biological activity in question are located. These are represented as virtual residues in a model of the active site. The method generates many different possible representations of the active site of the selected protein, presenting the user with a choice of premodeled molecules for synthesis and testing as a substitute for the protein.

Abstract: In one aspect of this invention, various protein/nucleic acid hybrid probes are described which can be used to amplify the detectable signal in immunoassays. The protein moiety is capable of functioning either as an antibody or an antigen. The nucleic acid moiety serves as a signal amplifier. In another aspect, various methods of amplifying the detectable signal in immunoassays by use of the hybrid probes and related polynucleotide probes are disclosed.

Abstract: Ribozymes designed to provide improved rates of catalytic turnover are described. The compounds of this invention comprise a catalytic region, at least one substrate binding region, and at least one displaceable antisense arm, whereby the rate of release of the endonuclease cleavage fragments is enhanced. A method to make such ribozymes is also described.

Abstract: A polydeoxynucleotide construct is disclosed for use, in conjunction with a DNA-dependent RNA polymerase, as a signal amplifier in nucleic acid hybridization assays. The construct contains a recognition sequence for a target oligonucleotide, a promoter sequence for a DNA-dependent RNA polymerase, and a polymerase template. A method of use for this construct in hybridization assays is also disclosed. The method involves formation of a hybridization complex comprising the construct and the target sequence; addition of a polymerase which is specific for the promoter in the construct; and quantification of the resulting RNA transcripts.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: The invention describes an assay device and assembly for detecting an analyte in a liquid sample. Each assay device in the assembly includes structure defining a well, a ligand-coated particle, and a flexible particle retaining structure for holding the particle in a captured position within the well.

Abstract: Novel DNA probe sequences for detection of Chlamydiae in a sample in a solution phase sandwich hybridization assay are described. Amplified nucleic acid hybridization assays using the probes are exemplified.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: A cover effective to releasably seal a multiwell container, such as a microtitration plate, is disclosed. The cover contains a pad, fashioned from a flexible polymer sheet, and a plurality of resiliently compressible ridges formed on the sheet. The ridges are deformable, such that application of pressure applied to the cover is effective to form a fluid-tight seal between the pad and the well openings. The ridges extend from the pad sufficiently to break the seal upon release of the pressure.

Abstract: The protease necessary for polyprotein processing in Hepatitis C virus is identified, cloned, and expressed. Proteases, truncated protease, and altered proteases are disclosed which are useful for cleavage of specific polypeptides, and for assay and design of antiviral agents specific for HCV.

Abstract: Methods and reagents are provided for synthesizing polynucleotides containing modified deoxyribose residues. Monomeric reagents having the structural formula (I) ##STR1## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are as defined herein, are used to create polynucleotides having nonnucleotidic moieties -A-Z-(R.sup.9).sub.n at the 1 position of selected deoxyribose units. The polynucleotides so provided are useful in a variety of hybridization assay formats.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: Methods and reagents are provided for synthesizing polynucleotides containing modified deoxyribose residues. Monomeric reagents having the structural formula (I) ##STR1## wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are as defined herein, are used to create polynucleotides having nonnucleotidic moieties --A--Z--(R.sup.9).sub.n at the 1 position of selected deoxyribose units. The polynucleotides so provided are useful in a variety of hybridization assay formats.

Abstract: Novel oligonucleotide probes are provided having the structure ##STR1## wherein: R.sup.2 is lower alkyl; R.sup.3 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 -- wherein R.sup.12 is hydrogen or lower alkyl; R.sup.5 is hydrogen or lower alkyl; R.sup.6 is selected from the group consisting of hydrogen, methyl, bromo and iodo; R.sup.7 is C.sub.1 -C.sub.12 alkylene containing 0 to 6 linkages selected from the group consisting of --O--, --S-- and --NR.sup.12 --, and bound to R.sup.8 through an --O--, --S-- or --NR.sup.12 -- moiety; and R.sup.8 is a protecting group that can be removed and replaced, without affecting the remainder of the compound, by reduction with a reducing agent. Methods for using the probes are provided as well.

Abstract: The protease necessary for polyprotein processing in Hepatitis C virus is identified, cloned, and expressed. Proteases, truncated protease, and altered proteases are disclosed which are useful for cleavage of specific polypeptides, and for assay and design of antiviral agents specific for HCV.

Abstract: Linear or branched oligonucleotide multimers useful as amplifiers in biochemical assays which comprise (1) at least one first single-stranded oligonucleotide unit that is complementary to a single-stranded oligonucleotide sequence of interest, and (2) a multiplicity of second single-stranded oligonucleotide units that are complementary to a single-stranded labeled oligonucleotide. Amplified sandwich nucleic acid hybridizations and immunoassays using the multimers are exemplified.

Abstract: A polynucleotide which has been modified by a hydrophobic moiety so that the polynucleotide attaches to a substrate by the hydrophobic moiety, rather than by a portion of the polynucleotide, thereby allowing the polynucleotide to be available for hybridization.

Abstract: Novel reagents useful in a variety of biochemical and chemical contexts, including nucleic hybridization assays and chemical phosphorylation of hydroxyl-containing compounds. The reagents are particularly useful for introducing cleavable sites and/or abasic sites into oligonucleotide or polynucleotide chains.

Abstract: Novel DNA probe sequences for detection of CMV in a sample in a solution phase sandwich hybridization assay are described. Amplified nucleic acid hybridization assays using the probes are exemplified.