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OwnerF. Gary Toback, John C. Lieskeinventors
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An autocrine crystal adhesion inhibitor called CAI is an anionic, sialic acid-containing glycoprotein secreted by kidney epithelial cells that blocks adhesion of calcium oxalate monohydrate (COM) crystals to the cell surfaces. Novel amino acid sequences are shown for the amino-acid terminus and 6 interval fragments. Persons may be classified according to risk of developing kidney stones, by measuring the amount of CAI in a biological sample. Treatment efficacy is also monitored by this method. CAI is administered in vivo to prevent nephrolithiasis. A rapid, simple assay to detect agents that inhibit adhesion of COM crystals to the surface of kidney epithelial cells is characterized.
A peptide having an amino acid sequence selected from the group consisting of the fifteen sequences as follows:
A method for determining if a candidate agent inhibits urinary crystal adhesion to kidney epithelial cells, said method comprising;
A method for screening a candidate agent for effectiveness in preventing kidney stone formation, said method comprising determining whether the agent inhibits crystal adhesion to cells.
The method of claim 8, wherein a tissue culture system comprising kidney epithelial cells is contacted with urinary crystals and the candidate agent, and the amount of crystals bound to the cells is compared with control cultures that are not contacted with the agent.
The method of claim 9, wherein the amount of crystals bound is compared to the amount of crystals bound when the renal epithelial cells are contacted with urinary crystals and the crystal adhesion inhibitor of claim 1
A quantitative method for monitoring a person at risk for kidney stone disease, said method comprising using a polyclonal antibody directed against the crystal adhesion inhibitor (CAI) to measure by an immunoassay the amount of CAI in a sample of urine from the person and comparing the amount so determined with that observed in persons that do not form kidney stones.
A method for classifying a kidney stone-former with respect to susceptibility to kidney stone formation, said method comprising comparing the activity of urinary CAI in a sample obtained from the stone-forming individual with samples from control individuals, said control individuals not being stoneformers.
The antibody of claim 14, further defined as directed to an active site of the purified inhibitor of claim 1.
The antibody of claim 14, further defined as a monospecific antibody.
The antibody of claim 16, further defined as a monoclonal antibody.
A monospecific antibody directed to the amino acid sequence NH2-Lys-Ile-Asn-Gly-Gly-Gly-Ala-Thr-Leu-Pro-Gln-Pro-Leu-Tyr-Gln-Thr-COOH.
A monospecific antibody directed to the amino acid sequence NH2-Leu-Asn-Asn-Asp-Tyr-Ser-Gln-Phe-Gly-Thr-Gly-Thr-Lys-COOH.
A cDNA sequence encoding a peptide comprising an amino acid sequence selected from the group consisting of the fifteen sequences as follows:
A cDNA molecule comprising, the following nucleotide sequence:
Dependencies are inferred from the claim text.