Abstract: A method is provided of making an oligonucleotide which comprises hybridizing (a) a shorter fragment of the desired oligonucleotide with (b) a nucleic acid fragment longer than (a) and complementary to the desired oligonucleotide, one of (a) and (b) is linked to an organic radical which differentiates its physical properties relative to the other of (a) and (b), contacting the hybridized material with an enzyme and nucleoside triphosphates whereby the shorter fragment is extended in one direction until it is substantially coterminal with the complementary nucleic acid fragment, denaturing the hybridized material and separating lengthened (a) from (b).

Abstract: A nucleic acid probe capable of participating in a chemiluminescent reaction comprising a defined nucleic acid sequence, the sequence being linked to any one ofa. a chemiluminescence precursor,b. a chemiluminescence enhancer, andc. an enzyme the remaining two of (a), (b) and (c) not linked to the sequence being in a mixture of the linked sequence. A method for determining a particular single stranded polynucleotide sequence in a test medium, comprising the steps of:(a) combining the test medium with a polynucleotide probe having a base sequence substantially complementary to the sequence to be determined,(b) labeling either the resulting hybrids or probe which has not hybridized with the sequence to be determined with one of the participants in an enhanced chamiluminescent reaction involving a chemiluminescent precursor, an enzyme, an oxidant, and a chemiluminescence enhancer,(c) initiating such chemiluminent reaction with the labeled hybrid or probe, and(d) detecting the resulting light emission.

Abstract: A nucleic acid detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion preferably including a recognition site for binding by a particular protein. Such recognition site can be a region of singly or doubly stranded nucleic acid specific for a particular nucleic acid binding protein such as lac repressor protein or can be a modified nucleic acid region such as a unique antigenic determinant introduced by interaction of the region with a modifier compound such as an intercalating agent or a platinum-containing ligand. The probe-binding protein can be labeled for ease of detection and in the case of an antigenic determinant binding site can be labeled antibody.

Abstract: A protein is covalently coupled to a 3'terminal end of a nucleic acid which carries several labels. In an assay the protein will specifically recognize some component of a test system; in an immunoassay the protein can be Protein A which will recognize the FC portion of IgG which is bound to an unknown antigen if present in the test sample.

Abstract: A labeled nucleic acid probe comprising (a) a nucleic acid component, (b) a nucleic acid-binding ligand photochemically linked to the nucleic acid component, and (c) a label chemically linked to the nucleic acid-binding ligand. The label can be a specifically bindable ligand such as a hapten or biotin, an enzyme such as a .beta.-galactosidase or horse radish peroxidase, a fluorescent radical, a phycobiliprotein, a luminescent radical, or a radioisotope. The probe can be used in assays of nucleic acids, taking advantage of the ability of the nucleic acid component to hybridize.

Abstract: A process for production of a single strand of a nucleic acid comprising covalently linking to a solid substrate a polynucleotide complementary to the desired strand, hybridizing said polynucleotide with an oligonucleotide, extending the oligonucleotide in direction away from said substrate, denaturing the hybridized polynucleotide and extended oligonucleotide, thereby to free the extended oligonucleotide from the solid substrate, and separating the extended oligonucleotide. The product can be used for making analytical probes.

Abstract: Monoclonal antibodies specific for the glucosylated N-terminal peptide residue in Hb A.sub.1c, a method for producing such antibodies, hybridoma cell lines secreting such antibodies and a method for their production, and immunoassay methods and reagent systems using such antibodies for the determination of Hb A.sub.1c in human blood samples. The monoclonal antibodies are secreted by hybridomas obtained from the fusion of a myeloma cell and a lymphocyte which has been taken from an animal, preferably a mouse, immunized with a synthetic peptide immunogen and which produces antibody specific for the glucosylated N-terminal peptide residue in Hb A.sub.1c. The synthetic peptide immunogen comprises an N-terminal plucosylated peptide residue having at least 2 amino acid units corresponding to the N-terminus of the beta-subunit of human hemoglobin and an immunogenic carrier to which the glucosylated peptide residue is linked.

Abstract: A detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion preferably including a recognition site for a particular protein.

Abstract: A solid support capable of binding a nucleic acid thereto upon suitable irradiation, comprising (a) a solid substrate, (b) a photochemically reactive intercalator compound or other nucleic acid-binding ligands, and (c) divalent radical chemically linking the substrate and the ligand (b). Specifically, a hydroxy group-containing solid substrate such as nitrocellulose paper is linked via a bifunctional reagent such as cyanogen bromide or 1,4-butanedioldiglycidyl ether to an amino-substituted angelicin or psoralen or ethidium bromide which in turn is photochemically linked to a nucleic acid. The resulting immobilized nucleic acid probe is capable of hybridizing with complementary nucleic acid fragments and is thereby useful in diagnostic assays.

Abstract: A substantially pure enzyme which hydrolyzes a polymer containing an .alpha. 2,8-linked N-acetyl neuraminic acid is obtained from K1-specific bateriophages and used for assays of bacterial polysaccharides in samples such as cerebrospinal fluid for testing for various disorders such as bacterial meningitis, septicemia, bacteremia, and the like. The enzyme may also be used therapeutically in treating such disorders by attacking cells which have an increased level of a polymer containing an .alpha. 2,8-linked N-acetyl neuraminic acid.

Abstract: A radioactively labeled protein comprising a protein, and a radioactive nucleoside or nucleotide, the protein being covalently linked to the nucleoside or nucleotide. Advantageously the linkage is through an NH.sub.2 group of the protein and through a carbonyl group of a ring-opened sugar moiety of the nucleoside or nucleotide. The protein can be insulin, an immunoglobulin or protein A. The radioactive moiety may be a P, C, S, H, I or Hg atom. The labels can be used to indicate the presence and amount of the protein in a biological assay.

Abstract: In a method for determining a particular polynucleotide sequence in a test medium containing single stranded nucleic acids wherein the sample is subjected to a hybridization reaction with a labeled detection probe having a substantially complementary polynucleotide sequence, and wherein after hybridization the label in said probe is assayed, the improvement wherein the label in said labeled probe comprises a fluorescent nucleotide which is linked by a phosphate ester linkage to said probe. Probes and kits therefor are also provided.

Abstract: Binding of a particular protein by an antibody reagent involving denaturation of the protein and use of an antibody reagent specific for binding a linear peptide epitope therein. Denaturation by chemical or physical means effectively exposes or enhances the exposure of the linear peptide epitope for binding by the antibody reagent which is preferably raised against a synthetic peptide immunogen. The technique is particularly useful in performing immunoassays for protein analytes, such as a glycosylated protein, in aqueous test samples.

Abstract: Monoclonal antibodies specific for the glucosylated N-terminal peptide residue in Hb A.sub.1c, a method for producing such antibodies, hybridoma cell lines secreting such antibodies and a method for their production, and immunoassay methods and reagent systems using such antibodies for the determination of Hb A.sub.1c in human blood samples. The monoclonal antibodies are secreted by hybridomas obtained from the fusion of a myeloma cell and a lymphocyte which has been taken from an animal, preferably a mouse, immunized with a synthetic peptide immunogen and which produces antibody specific for the glucosylated N-terminal peptide residue in Hb A.sub.1c. The synthetic peptide immunogen comprises an N-terminal glucosylated peptide residue having at least 2 amino acid units corresponding to the N-terminus of the beta-subunit of human hemoglobin and an immunogenic carrier to which the glucosylated peptide residue is linked.

Abstract: A solid support capable of binding a nucleic acid thereto upon suitable irradiation, comprising (a) a solid substrate, (b) a member selected from the group consisting of a furocoumarin, a phenanthridium halide, and photochemically reactive derivatives thereof, and (c) a divalent radical chemically linking the substrate and the member (b). Specifically, a hydroxy group-containing solid substrate such as nitrocellulose paper is linked via a bifunctional reagent such as cyanogen bromide or 1,4-butanediol diglycidyl ether to an amino-substituted angelicin or psoralen or phenanthridinium bromide which in turn is photochemically linked to a nucleic acid. This is capable of hybridizing with other nucleic acid fragments and is thereby useful in diagnostic assays.