Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
2001-1-26
pubmed:abstractText
A subclone of the NS0 murine myeloma cell line, frequently used to produce recombinant monoclonal antibodies, was found by a transmission electron microscopy method to express a surprisingly high titer of 10(11) retroviral particles per ml of culture supernatant. Infectivity assays showed a very low infectious titer with the restricted host range expected for a murine amphotropic retrovirus. A Western blot assay for the viral capsid protein was developed to confirm the high titer values and provide a means for monitoring batch consistency and virus removal during the purification process. Mass spectrometry of several of the viral Gag proteins demonstrated that the cell line appeared to produce at least two closely related retroviruses. N-terminal sequencing of three of the Gag proteins demonstrated that these retroviruses were members of the murine leukemia retroviral family. Western blot detection with an antibody for the capsid protein gave a linear standard curve over the range of 0.1-3 ng per lane. This allows the detection of viral titers as low as 6x10(7) virions per ml without the need to concentrate the sample. The Western blot method has higher throughput and less variability than transmission electron microscopy methods and has potential for monitoring viral titer and clearance during development of manufacturing processes.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0168-1656
pubmed:author
pubmed:issnType
Print
pubmed:day
17
pubmed:volume
84
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
33-43
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11035185-Amino Acid Sequence, pubmed-meshheading:11035185-Animals, pubmed-meshheading:11035185-Bioreactors, pubmed-meshheading:11035185-Blotting, Western, pubmed-meshheading:11035185-Capsid, pubmed-meshheading:11035185-Chromatography, High Pressure Liquid, pubmed-meshheading:11035185-Gene Products, gag, pubmed-meshheading:11035185-Leukemia, Experimental, pubmed-meshheading:11035185-Leukemia Virus, Murine, pubmed-meshheading:11035185-Mass Spectrometry, pubmed-meshheading:11035185-Mice, pubmed-meshheading:11035185-Microscopy, Electron, pubmed-meshheading:11035185-Molecular Sequence Data, pubmed-meshheading:11035185-Multiple Myeloma, pubmed-meshheading:11035185-Retroviridae Infections, pubmed-meshheading:11035185-Retroviridae Proteins, pubmed-meshheading:11035185-Tumor Cells, Cultured, pubmed-meshheading:11035185-Tumor Virus Infections, pubmed-meshheading:11035185-Viral Load
pubmed:year
2001
pubmed:articleTitle
Biochemical analysis of retroviral structural proteins to identify and quantify retrovirus expressed by an NS0 murine myeloma cell line.
pubmed:affiliation
Biogen Inc., 14 Cambridge Center, Cambridge, MA 02142, USA. fred_taylor@biogen.com
pubmed:publicationType
Journal Article