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Effect of Protease Inhibitors on Yield of HSV‐1‐Based Viral Vectors
Author(s) -
Wechuck James B.,
Goins William F.,
Glorioso Joseph C.,
Ataai Mohammad M.
Publication year - 2000
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp000016t
Subject(s) - protease , virology , vector (molecular biology) , protease inhibitor (pharmacology) , titer , herpes simplex virus , hsl and hsv , yield (engineering) , biology , recombinant dna , chemistry , virus , enzyme , biochemistry , gene , viral load , physics , antiretroviral therapy , thermodynamics
The ability to obtain high titer replication‐defective herpes simplex virus (HSV) recombinant vectors will dramatically affect their use in gene therapy clinical trials. A variety of techniques and reagents have been employed to increase the overall yield of the vector. The effects of protease inhibitors on the yield of an HSV‐1‐based viral vector were examined. Experiments were conducted using a commercial protease inhibitor cocktail typically used in mammalian cell culture for protein production. Contrary to our expectation for enhanced vector yield, the results showed a dramatic reduction in vector yield. Moreover, it was found that AEBSF is the only component in the protease cocktail responsible for the low vector yield. On the basis of our hypothesis regarding the mode of action of AEBSF, we suggest that it should not be included in protease inhibitor cocktails designed for use in cultures aimed at production of viral vectors derived from HSV‐1 or possibly several other vectors.