US2003170710A1PendingUtilityA1

Method of separating viral particles

46
Assignee: AVENTIS PHARMA SAPriority: Dec 31, 1998Filed: Feb 25, 2003Published: Sep 11, 2003
Est. expiryDec 31, 2018(expired)· nominal 20-yr term from priority
Inventors:Francis BlancheAnne BarbotBeatrice Cameron
G01N 30/02G01N 2333/075G01N 33/56983G01N 33/538C12Q 1/24C12N 2710/10351C12N 7/00C12N 7/02
46
PatentIndex Score
0
Cited by
0
References
0
Claims

Abstract

The invention concerns a novel method for purifying and quantifying viral particles. More particularly, the invention concerns a method for purifying and quantifying adenovirus by ion-exchange chromatography. The invention also concerns a method for identifying different adenovirus serotypes.

Claims

exact text as granted — not AI-modified
1 . Method of separating viral particles from a biological medium, characterized in that it comprises at least one chromatography step carried out on a support comprising a matrix and ion-exchange groups, the said groups being grafted onto the said matrix by means of a flexible arm.  
     
     
         2 . Method according to  claim 1 , characterized in that the matrix is chosen from agarose, dextran, acrylamide, silica and poly[styrene-divinylbenzene], alone or in the form of a mixture.  
     
     
         3 . Method according to  claim 2 , characterized in that the matrix consists of cross-linked agarose and preferably 6% cross-linked agarose.  
     
     
         4 . Method according to one of  claims 1  to  3 , characterized in that the matrix has a particle size of between about 40 and 200 μm.  
     
     
         5 . Method according to  claim 4 , characterized in that the matrix has a particle size of between 45 and 165 μm and is centered on 90 μm.  
     
     
         6 . Method according to  claim 4  or  5 , characterized in that the matrix has a dispersion such that 95% of the particles have a diameter of between 0.1 and 10 times the mean diameter of the particles, and preferably between 0.3 and 3 times the mean diameter of the particles.  
     
     
         7 . Method according to  claim 1 , characterized in that the flexible arm is a hydrophilic arm consisting of a polymer of synthetic or natural origin.  
     
     
         8 . Method according to  claim 7 , characterized in that the flexible arm is a polymer of synthetic origin chosen from polyvinyl alcohols, polyacrylamides, polymethacrylamides or polyvinyl ethers.  
     
     
         9 . Method according to  claim 7 , characterized in that the flexible arm is a polymer of natural origin of a polysaccharide nature chosen from starch, cellulose, dextran and agarose.  
     
     
         10 . Method according to  claim 8  or  9 , characterized in that the degree of polymerization of the flexible arm is about 30 monomeric units.  
     
     
         11 . Method according to  claim 10 , characterized in that the flexible arm is a dextran having a mean molecular weight of about 5000 Da.  
     
     
         12 . Method according to  claim 1 , characterized in that the ion-exchange group is a strong anion-exchange group.  
     
     
         13 . Method according to  claim 12 , characterized in that the strong anion-exchange group is a quaternary amine.  
     
     
         14 . Method according to one of  claims 7  to  13  or according to  claim 5 , characterized in that the chromatography is carried out on a Q Sepharose® XL type support.  
     
     
         15 . Method according to  claim 1 , characterized in that the biological medium is a supernatant of encapsidation cells producing the said virus.  
     
     
         16 . Method according to  claim 1 , characterized in that the biological medium is a lysate of encapsidation cells producing the said virus.  
     
     
         17 . Method according to  claim 1 , characterized in that the biological medium is a prepurified solution of the said virus.  
     
     
         18 . Method according to  claim 1 , characterized in that it comprises a preliminary ultrafiltration step.  
     
     
         19 . Method according to  claim 18 , characterized in that the ultrafiltration is a tangential ultrafiltration on a membrane having a cut-off of between 300 and 500 kDa.  
     
     
         20 . Use of a Q Sepharose® XL type chromatography support for the analytical and/or preparative separation of viral particles.  
     
     
         21 . Use according to  claim 20 , characterized in that the viral particles are adenoviruses.  
     
     
         22 . Use of a Q Sepharose® XL type chromatography support for the identification of various adenovirus serotypes.  
     
     
         23 . Use of a Q Sepharose® XL type chromatography support for the titration of adenoviruses.  
     
     
         24 . Method of quantifying adenoviruses, characterized in that the viral particles are separated by chromatography on a Q Sepharose® XL type support and the quantity of adenoviruses is measured by the absorbance of the chromatographic fractions.

Join the waitlist — get patent alerts

Track US2003170710A1 — get alerts on status changes and closely related new filings.

We store only your email — no account needed. See our privacy policy.