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Bioreactor Production of Recombinant Herpes Simplex Virus Vectors
Author(s) -
Knop David R.,
Harrell Heather
Publication year - 2008
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/bp060373p
Subject(s) - herpes simplex virus , vector (molecular biology) , bioreactor , virology , biology , virus , viral vector , recombinant dna , shuttle vector , gene , genetics , botany
Abstract Serotypical application of herpes simplex virus (HSV) vectors to gene therapy (type 1) and prophylactic vaccines (types 1 and 2) has garnered substantial clinical interest recently. HSV vectors and amplicons have also been employed as helper virus constructs for manufacture of the dependovirus adeno‐associated virus (AAV). Large quantities of infectious HSV stocks are requisite for these therapeutic applications, requiring a scalable vector manufacturing and processing platform comprised of unit operations which accommodate the fragility of HSV. In this study, production of a replication deficient rHSV‐1 vector bearing the rep and cap genes of AAV‐2 (denoted rHSV‐rep2/cap2) was investigated. Adaptation of rHSV production from T225 flasks to a packed bed, fed‐batch bioreactor permitted an 1100‐fold increment in total vector production without a decrease in specific vector yield (pfu/cell). The fed‐batch bioreactor system afforded a rHSV‐rep2/cap2 vector recovery of 2.8 × 10 12 pfu. The recovered vector was concentrated by tangential flow filtration (TFF), permitting vector stocks to be formulated at greater than 1.5 × 10 9 pfu/mL.