Biomedical Engineering Reference
In-Depth Information
30.
Biotechnol J
. 2008 Oct; 3(9-
10):1185-200. Implementation of
advanced technologies in
commercial monoclonal
antibody production. Zhou JX,
Tressel T, Yang X, Seewoester T.
Process and Product
Development, Amgen, Inc.,
Thousand Oaks, CA 91320-1799.
Process advancements driven through innovations
have been key factors that enabled successful
commercialization of several human therapeutic
antibodies in recent years. The production costs of
these molecules are higher in comparison to
traditional medicines. In order to lower the
development and later manufacturing costs, recent
advances in antibody production technologies target
higher-throughput processes with increased clinical
and commercial economics. In this review, essential
considerations and trends for commercial process
development and optimization are described,
followed by the challenges to obtain a high titer cell
culture process and its subsequent impact on the
purification process. One of these recent technical
advances is the development and implementation of
a disposable Q membrane adsorber as an alternative
to a Q-packed-bed column in a flow-through mode.
The scientific concept and principles underlining Q
membrane technology and its application are also
reviewed.
31.
J Biotechnol
. 2008 Jun 30;
135(3):272-80. Epub 2008 May
21. Expression of SEAP (secreted
alkaline phosphatase) by
baculovirus mediated
transduction of HEK 293 cells in
a hollow fiber bioreactor system.
Jardin BA, Zhao Y, Selvaraj M,
Montes J, Tran R, Prakash S,
Elias CB. Biotchnology Research
Institute, 6100 Royalmount
Avenue, Montreal, Quebec,
Canada.
A BacMam baculovirus was designed in our
laboratory to express the reporter protein secreted
alkaline phosphatase (SEAP) driven by the
immediate early promoter of human
cytomegalovirus promoter (CMV). In vitro tests have
been carried out using this recombinant baculovirus
to study the secreted protein in two cell lines and
under various culture conditions. The transductions
were carried out on two commonly used mammalian
cell lines, namely, the human embryonic kidney
(HEK 293A) and Chinese hamster ovary (CHO-K1).
Initial studies clearly demonstrated that the transient
expression of SEAP was at least tenfold higher in the
HEK 293 cells than the CHO cells under equivalent
experimental conditions. Factorial design
experiments were done to study the effect of
different parameters such as cell density, multiplicity
of infection (MOI), and the histone deacetylase
inhibitor, trichostatin A concentration. The MOI and
the cell density were found to have the most impact
on the process. The enhancer trichostatin A also
showed some positive effect. The production of
secreted protein in a batch reactor was studied using
the Wave disposable bioreactor system. A
semicontinuous perfusion process was developed to
extend the period of gene expression in mammalian
cells using a hollow fiber bioreactor system (HFBR).
The growth of cells and viability in both systems was
monitored by offline analyses of metabolites. The
expression of recombinant protein could be
maintained over an extended period of time up to 30
days in the HFBR.
