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"prionins", highly specific markers for noninvasive pre-symptomatic detection of tse diseases, and targets for therapeutic reagents to prevent and control tse diseases in animals and humans

a tse disease and highly specific technology, applied in the field of prionins, can solve the problems of unsatisfactory explanation and unsubstantial progress in these directions

Inactive Publication Date: 2002-10-24
BERGMANN JOHANNA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0019] In general, first we identified a gene closely related to a gene already genetically or otherwise linked to a certain disease, then isolated the mRNA transcribed from the gene from disease tissue or patient's blood, then synthesized cDNA from the isolated mRNA with reverse transcriptase, then amplified the novel cDNA with specific primers which flanked the entire coding region of the cDNA, then we identified the cDNA from the size following electrophoresis on agarose gel, and finally isolated the unique cDNA from the agarose gel. This allowed us to select out the desired molecule, if it was expressed, without having to probe several million cDNA clones.

Problems solved by technology

Nevertheless, it has not been satisfactorily explained how a brain resident protein which, so far has not been demonstrated in biological fluids, can be transmitted from animal to animal within a grazing herd of cattle.
Nevertheless despite a large international effort to understand the pathogenesis of TSE, and to develop non invasive widely applicable methods for presymptomatic detection of these diseases, which in the case of BSE and scrapie have calamitous effects on commercial activity and on the health of the population at large, substantial progress has not been made in these directions.

Method used

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  • "prionins", highly specific markers for noninvasive pre-symptomatic detection of tse diseases, and targets for therapeutic reagents to prevent and control tse diseases in animals and humans
  • "prionins", highly specific markers for noninvasive pre-symptomatic detection of tse diseases, and targets for therapeutic reagents to prevent and control tse diseases in animals and humans
  • "prionins", highly specific markers for noninvasive pre-symptomatic detection of tse diseases, and targets for therapeutic reagents to prevent and control tse diseases in animals and humans

Examples

Experimental program
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example 2

[0026] Detection of BSAS, SCRAPAS and CJAS in clinical and other samples.

[0027] Prionin proteins were detected using the ELISAs described above. Since some populations ft the polyclonal anti-TSE antibodies cross react with domains of the major epitopes of each TSE species, with varying degrees of sensitivity, anti-SCRAPAS-HRP is used as the second antibody for detecting both BSAS and SCHAPAS, whereas either anti-SCRAPAS HRP or anti-CJAS-HRP is used as the second antibody for detecting CJAS.

example 3

[0028] Using another approach, which is not suitable for use as a routine test method (with BSAS as the example) : BSAS protein was isolated from 100 .mu.l of serum, from an infected cow on a anti-BSAS antibody immuno-affinity column. The isolated protein was subjected to SDS gel electrophoresis as described by Schgger, H. and von Jabow, G. I. Anal. Biochem. 166:368-379 (1987) or by non-SDS PAGE. Following electrophoresis the proteins were subjected to Western blotting or spotted onto nylon membrane and treated with the affinity purified antibody. Interaction of the antibody with the protein bound to the membrane was visualized with a chemiluminescent kit purchased from Bio-Rad Inc. according to the manufacturer's instructions (also see Blake M. S. et al.Anal biochem. 136:175-179 (1984)). MOPAS and HAMPAS were discovered using the same procedure; however, the expression of these two proteins in animals with experimental TSE disease have not been investigated so far.

example 4

[0029] We have demonstrated, in the case of BSAS and SCHAPAS, a specific association (100%) of the proteins with animals confirmed with the disease or exposed in any manner to the disease; whereas the proteins were not associated with animals which have never been exposed to the disease. BSAS and SCHAPAS was detected in serum of all animals clinically positive for the diseases and in the majority of animal that had no demonstrated clinical symptoms of the disease which came from herds that had even a single case of the disease. CJAS was detected in serum of two CJD victims in one of which was positive for both CJD and Alzheimer's disease.

[0030] The success of the present invention derived from our ability to find alternative genes encoding potentially pathogenic proteins BSAS, SCRAPAS and CJAS within the PrP genes of cattle, sheep and humans. The discovery of equivalent genes in mice and hamsters contributed to formulating our model for TSE.

Details of the Invention

[0031] In detail t...

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Abstract

Proteins expressed from within the prion protein genes of all animals and humans, "prionins", against which reagents can be prepared for accurate pre-symptomatic diagnosis, for detecting latent TSE, for detecting TSE contamination of food, blood and blood products and for therapeutic treatment of Bovine spongiform encephalopathy (BSE) in cows, Scrapie disease in sheep and Creutzfeldt-Jakob syndrome in humans, are revealed.

Description

[0001] This invention is related to diagnostic and therapeutic molecules for the detection, prevention of bovine spongiform encephalopathy (BSE, scrapie disease (scrapie) and Creutzfeldt-Jakob syndrome (CJS). Specifically, the invention relates to three closely related Proteins BSAS, SCRAPAS and CJAS which are implicated in causing BSE, scrapie and CJS, to antagonists of these proteins, to diagnostic reagents to detect these proteins in clinical samples and food, and to therapeutic methods directed at these proteins in animals and humans. It is also related to the use of homologues of these proteins from hamsters and mice which are useful for developing and testing methods for use with vaccines and other agents for therapeutic value in treatment of humans and animals for TSE diseases.[0002] Prion proteins (PrPs) are a family of very closely related proteins which are found in a number of allelic forms in the membrane of certain populations of brain neurons of all animals. PrPs are e...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395A61P25/28C07K14/47C07K16/18C12N15/09C12Q1/68G01N33/53G01N33/68
CPCC07K14/47G01N2800/2828G01N33/6896C07K16/18A61P25/28Y10S435/962Y10S435/97
Inventor BERGMANN, JOHANNAPREDDIE, ENRIQUE
Owner BERGMANN JOHANNA
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