Document Detail


A simple methodology for the routine production and partial purification of human lymphoblastoid interferon.
MedLine Citation:
PMID:  6328906     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We report a methodology suitable for the large scale production and partial purification of human lymphoblastoid interferon with a minimum expense and reasonable (10%) degree of purity of product. The cells used were Namalwa lymphoblastoid cells, which have the advantage of being both easy to grow and well characterized. They were grown in RPMI 1640 containing 10% fetal calf serum to a density of 1.5 - 2 X 10(6) cells/ml and then diluted 50% by the addition of serum free medium containing 2mM sodium butyrate. After 48 hours, the medium was removed and the cells induced to produce interferon by Sendai virus. Typical initial interferon titres were in the region of 4 - 4.8 log units/ml, while specific activities were in the region of 10(6) units/mg protein. The initial stage in the purification involved batchwise adsorption of the interferon to Procion red- HE7B Sepharose CL6B . The Sepharose was then packed into a column, washed with 0.5M KC1 and the interferon eluted with 2M KC1. The interferon was further purified by gel filtration to give an activity of approximately 10(7) units/mg protein. Yields were between 30-50% of the initial interferon in a volume of 25 ml. Further separation of the components of the heterogeneous alpha interferon could be obtained on Procion blue-HER Sepharose, or by utilizing reverse phase HPLC.
Authors:
P J Neame; R T Acton
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Advances in experimental medicine and biology     Volume:  172     ISSN:  0065-2598     ISO Abbreviation:  Adv. Exp. Med. Biol.     Publication Date:  1984  
Date Detail:
Created Date:  1984-07-02     Completed Date:  1984-07-02     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0121103     Medline TA:  Adv Exp Med Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  269-79     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Burkitt Lymphoma
Cell Line
Cercopithecus aethiops
Humans
Interferon Type I / biosynthesis*,  isolation & purification,  toxicity
Kidney
Parainfluenza Virus 1, Human
Semliki forest virus / drug effects
Grant Support
ID/Acronym/Agency:
CA09128/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Interferon Type I

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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