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Tumour-Specific Animal Proteins

a technology of animal proteins and tumors, applied in the field of tumor-specific animal proteins, can solve the problems of autoimmune induction and inappropriate immune respons

Inactive Publication Date: 2010-11-18
GLAXOSMITHKLINE BIOLOGICALS SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, their inappropriate expression in certain circumstances can cause an induction of autoimmune, inappropriate immune responses, which could be corrected through appropriate vaccination using the same polypeptides or polynucleotides.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0253]Real-Time RT-PCR Analysis in Matched Tumour and Normal Colon Tissues from Multiple Patients

[0254]Real-time RT-PCR (U. Gibson. 1996. Genome Research: 6,996) is used to compare mRNA transcript abundance of the candidate antigen in matched tumour and normal colon tissues from multiple patients. In addition, mRNA levels of the candidate gene in a panel of normal tissues are also evaluated by this approach.

[0255]Total RNA from normal and tumour colon is extracted from snap frozen biopsies using TriPure reagent (Boehringer). Total RNA from normal tissues is purchased from InVitrogen or is extracted from snap frozen biopsies using TriPure reagent. Poly-A+ mRNA is purified from total RNA after DNAase treatment using oligo-dT magnetic beads (Dynal). Quantification of the mRNA is performed by spectrofluorimetry (VersaFluor, BioRad) using SybrIl dye (Molecular Probes). Primers for real-time PCR amplification are designed with the Perkin-Elmer Primer Express software using default options...

example 2

[0262]Differential Screening of cDNA Arrays

[0263]Identification of tumour-associated genes in the subtracted cDNA library is accomplished by differential screening.

[0264]Total bacterial DNA is extracted from 100 μl over-night cultures. Bacteria are lysed with guanidium isothiocyantate and the bacterial DNA is affinity purified using magnetic glass (Boehringer). Plasmid inserts are recovered from the bacterial DNA by Advantage PCR amplification (Clontech). The PCR products are dotted onto two nylon membranes to produce high density cDNA arrays using the Biomek 96 HDRT tool (Beekman). The spotted cDNA is covalently linked to the membrane by UV irradiation. The first membrane is hybridised with a mixed cDNA probe prepared from the tumour of a single patient. The second membrane is hybridised with an equivalent amount of mixed cDNA probe prepared from normal colon of the same patient. The probe cDNA is prepared by PCR amplification as described above and is labelled using the AlkPhos Di...

example 3

DNA Microarrays

[0266]DNA micro-arrays are used to examine mRNA expression profiles of large collections of genes in multiple samples. This information is used to complement the data obtained by real-time PCR and provides an independent measure of gene expression levels in tumors and normal tissues.

[0267]Examples of current technologies for production of DNA micro-arrays include 1) The Affymetrix “GeneChip” arrays in which oligonucleotides are synthetized on the surface of the chip by solid phase chemical synthesis using a photolithographic process 2) DNA spotting technology in which small volumes of a DNA solution are robotically deposited and then immobilized onto the surface of a solid phase (e.g. glass). In both instances, the chips are hybridized with cDNA or cRNA which has been extracted from the tissue of interest (e.g. normal tissue, tumour etc . . . ) and labeled with radioactivity or with a fluorescent reporter molecule. The labeled material is hybridized to the chip and th...

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Abstract

CASB7439 polypeptides and polynucleotides, immunogenic compositions comprising them and methods for producing such polypeptides by recombinant techniques are disclosed. Also disclosed are methods for utilizing CASB7439 polypeptides and polynucleotides in diagnostics, and vaccines for prophylactic and therapeutic treatment of cancers, particularly colorectal, breast, and lung cancers, autoimmune diseases, and related conditions.

Description

CROSS REFERENCES TO RELATED APPLICATIONS[0001]This application is a continuation-in-part and claims benefit to U.S. application Ser. No. 10 / 226,872, which was filed in the United States Patent and Trademark Office on Aug. 23, 2002, which was filed as a continuation-in-part of International Application No. PCT / EP01 / 01779, filed on Feb. 16, 2001, which claims priority of Great Britain Patent Application No. 0004269.7, filed Feb. 23, 2000, which claims priority of Great Britain Patent Application No. 0009905.1; filed Apr. 20, 2000, which claims priority of Great Britain Patent Application No. 0021080.7, filed Aug. 25, 2000. This application also claims benefit to copending International Application Serial Nos.: PCT / EP02 / 05011, filed 16 May 2001 and PCT / EP02 / 01649, filed 21 Feb. 2001, the contents of which are incorporated herein by reference in their entirety.FIELD OF THE INVENTION[0002]The present invention relates to pharmaceutical compositions and methods for inducing an immune resp...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/00A61P37/04A61K35/12G01N33/50A61K38/00A61K39/39A61K45/00A61K48/00A61P1/00A61P35/00C07K14/47C07K14/82C07K16/18C12N1/15C12N1/19C12N1/21C12N5/10C12N15/09C12P21/02C12Q1/68G01N33/15G01N33/68
CPCA61K38/00C07K14/82C07K14/4748A61K39/00A61P1/00A61P35/00A61P37/02A61P37/04C07K14/47
Inventor CASSART, JEAN-POLCABEZON-SILVA, TERESA ELISA VIRGINIACOCHE, THIERRYGAULIS, SWANN ROMAIN JEAN-THOMASBASSOLS, CARLOTA VINALS Y DE
Owner GLAXOSMITHKLINE BIOLOGICALS SA
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