Mammalian proteinases; oxidoreductases; related reagents

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...

Reexamination Certificate

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C530S388100, C530S388260, C530S389100, C530S391300, C435S975000, C435S007100, C424S146100, C424S139100, C424S130100

Reexamination Certificate

active

06518405

ABSTRACT:

FIELD OF THE INVENTION
The present invention contemplates compositions related to proteins from animals, e.g., mammals, which function as proteinases; or oxidoreductases. In particular, it provides nucleic acids which encode, antibodies to, and proteins which exhibit biological functions, e.g., capacity to degrade proteinaceous substrates or serve as oxidoreductases.
BACKGROUND OF THE INVENTION
The proteases are a very broad group of enzymes which carry out an enzymatic function of hydrolysing a peptide bond. Within the group, there is a wide range of substrate specificities for the amino acids adjacent the cleavage sites. Proteases are typically categorized on the basis of their catalytic mecahnisms, e.g., based upon studies of their active sites, or by the effects of pH. Four main categories of proteases are serine proteinases, sulfhydryl proteases, acid proteases, and metalloproteases. They may also be classified according to their cleavage sites, e.g., endoproteases, amino peptidases, or carboxy peptidases.
Proteases have traditionally held a large share of the industrial enzyme market. Proteases are used in many industrial processes, including in detergents and cleaning products, e.g., to degrade protein materials such as blood and stains, in leather production, e.g., to remove hair, in baking, e.g., to break down glutens, in flavorings, e.g., soy sauce, in meat tenderizing, e.g., to break down collagen, in gelatin or food supplement production, in the textile industry, in waste treatment, and in the photographic industry. See, e.g., Gusek (1991)
Inform
1:14-18; Zamost, et al. (1996)
J. Industrial Microbiol.
8:71-82; James and Simpson (1996)
CRC Critical Reviews in Food Science and Nutrition
36:437-463; Teichgraeber, et al. (1993)
Trends in Food Science and Technology
4:145-149; Tjwan, et al. (1993)
J. Dairy Research
60:269-286; Haard (1992)
J. Aquatic Food Product Technology
1:17-35; van Dijk (1995)
Laundry and Cleaning News
21:32-33; Nolte, et al. (1996)
J. Textile Institute
87:212-226; Chikkodi, et al. (1995)
Textile Res. J.
65:564-569; and Shih (1993)
Poultry Science
72:1617-1620.
Oxidoreductases are involved in oxidation and reduction reactions, and have important functions, e.g., in oxidative phosphorylation. These enzymes are important in electron transport and general aerobic metabolism, and in many cases are associated with the mitochondrial membranes. In various circumstances, it may be useful to modulate oxidoreductase reactions to slow down or increase energy metabolism, e.g., in a cell or organ.
While there are many uses for proteases, there is always the need for a more active protease under various specific conditions. Similarly, regulation of oxidoreduction may be important. Alternatively, the distribution of these gene products may be useful as markers for specific cell or tissue types. There is a need for new proteinases or oxidoreductase enzymes of differing properties, specificities, and activities.
SUMMARY OF THE INVENTION
The present invention provides a binding compound comprising an antibody binding site which specifically binds to primate BS10.55 protein; or primate YTF03 protein; a nucleic acid comprising sequence encoding at least 12 amino acids of primate BS10.55 protein; or primate YTF03 protein; a substantially pure protein which is specifically recognized by the above antibody binding site; a substantially pure primate BS10.55, or primate YTF03 protein or peptide thereof; and a fusion protein comprising a 30 amino acid sequence portion of primate BS10. 55, or primate YTF03 protein sequence.
In certain binding compound embodiments, the antibody binding site is specifically immunoreactive with a protein selected from polypeptides of SEQ ID NO: 2 and 4; is raised against a purified or recombinantly produced primate BS10.55, or primate YTF3 protein; is immunoselected on a substantially purified or recombinantly produced primate BS10.55, or primate YTF03 protein; is in a monoclonal antibody, Fab, or F(ab)2; is detectably labeled; is attached to a solid substrate; is from a rabbit or mouse; binds with a Kd of at least about 300 mM; is fused to another protein segment; is in a chimeric antibody; or is coupled to another chemical moiety.
The invention also provides a method of making an antigen-antibody complex, comprising a step of contacting a primate biological sample to a specific binding antibody described. In preferred embodiments, the method further includes steps to purify the antigen or antibody.
Alternative embodiments provide an antibody binding site wherein the binding site is detected in a biological sample by a method comprising the steps of contacting a binding agent having an affinity for BS10.55 or YTF03 protein with the biological sample; incubating the binding agent with the biological sample to form a binding agent: BS10.55 or binding agent: YTF03 protein complex; and detecting the complex. In certain embodiments, the biological sample is human, and the binding agent is an antibody.
The invention also provides kits containing a composition described above and instructional material for the use of the composition; or segregation of the composition into a container. Typically, the kit is used to make a qualitative or quantitative analysis.
The invention also embraces a cell comprising an antibody described above; a cell transfected with a nucleic acid described above; or a cell comprising a fusion protein described above.
In nucleic acid embodiments, the nucleic acid may encode a polypeptide which specifically binds an antibody generated against an immunogen selected from the group consisting of the mature polypeptides of SEQ ID NO: 2 and 4. Alternatively, the nucleic acid may encode at least 12 amino acids of SEQ ID NO: 2 or SEQ ID NO: 4; comprise sequence of at least about 39 nucleotides selected from protein coding portions of SEQ ID NO: 1 or 3; hybridize to SEQ ID NO: 1 or 3 under stringent wash conditions of at least 45° C. and less than about 150 mM salt; comprise sequence made by a synthetic method; be an expression vector; be detectably labeled; be attached to a solid substrate; be from human; bind with a Kd of at least about 300 &mgr;M; be fused to another nucleic acid segment; be coupled to another chemical moiety; be operably associated with promoter, ribosome binding site, or poly-A addition site; be a PCR product; be transformed into a cell, including a bacterial cell; be in a sterile composition; be capable of selectively hybridizing to a nucleic acid encoding a BS10.55, or YTF03 protein; comprise a natural sequence; comprise a mature protein coding segment of SEQ ID NO: 1 or 3; encode proteolytically active portion of BS10.55; encode an oxidoreductive active portion of YTF03; be detected in a biological sample by a method comprising: contacting a biological sample with a nucleic acid probe capable of selectively hybridizing to said nucleic acid, incubating the nucleic acid probe with the biological sample to form a hybrid of the nucleic acid probe with complementary nucleic acid sequences present in the biological sample; and determining the extent of hybridization of the nucleic acid probe to the complementary nucleic acid sequences, including the method where the nucleic acid probe is capable of hybridizing to a nucleic acid encoding a protein selected from the group consisting of the mature polypeptides of SEQ ID NO 2 and 4.
In protein or polypeptide embodiments, the proteins may bind with a Kd of at least about 30 &mgr;M to an antibody generated against an immunogen of the polypeptides of SEQ ID NO: 2 or 4; be immunoselected on an antibody which selectively binds a polypeptide of SEQ ID NO: 2 or 4; comprise sequence of at least 12 contiguous residues of SEQ ID NO: 2 or 4; exhibit a post-translational modification pattern distinct from natural BS10.55, or YTF03; be 3-fold or fewer substituted from natural sequence; be recombinantly produced; be denatured; have sequence of full length natural polypeptide; be detectably labeled; be attached to a solid substrate; be from human; be in a sterile co

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