title: The use of UCOE vectors in combination with a preadapted serum free, suspension cell line allows for rapid production of large quantities of protein.
abstract: UCOE vectors contain non-tissue specific chromatin-opening-elements that permit rapid expression of a protein in anintegration independent manner. Efficient expression can bederived from a single copy of an integrated gene site resulting ina higher percentage of cells expressing the marker gene in theselected pool in comparison to standard non-UCOE containingvectors. This, in combination with the utilization of a serum-free, suspension adapted parent cell line allows for rapidproduction of large quantities of protein in a short period oftime. Utilizing this system more than 300 mg of a recombinantantibody has been produced in less than 1 month from transfectionpools in shake flask. Selected subclones have been scaled intosmall bioreactors in less than 2 months, producing significantquantities of monoclonal antibody using a protocol generic for theparent cell line. The increased efficiency obtained with the UCOEvector reduces the number of transfectants which need to bescreened in order to obtain high productivity subclones.Transfection of a standard host cell line, preadapted to grow in alarge-scale setting, allows for rapid cell line developmentdecreasing the transition time from research into development andmanufacturing. Alternatively, the traditional approach of using aparent cell line which requires serum-free and suspensionadaptation after transfection further increases the need forscreening a large number of subclones, because many of thesubclones will not be able to grow under conditions that allowlarge-scale protein production. The use of a preadapted cell linecan reduce the time required to develop a cell line from months toweeks.
authors.affiliation: Hyseq Pharmaceuticals, 675 Almanor Ave., Sunnyvale, CA, 94086, USA.
authors.forename: Trish
authors.initials: T
authors.lastname: Benton
authors.forename: Tim
authors.initials: T
authors.lastname: Chen
authors.forename: Michele
authors.initials: M
authors.lastname: McEntee
authors.forename: Brian
authors.initials: B
authors.lastname: Fox
authors.forename: David
authors.initials: D
authors.lastname: King
authors.forename: Robert
authors.initials: R
authors.lastname: Crombie
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Thomas
authors.forename: Christopher
authors.initials: C
authors.lastname: Bebbington