Advanced Granulation Technology™ (AGT™) Media for ... - Invitrogen

Advanced Granulation Technology
(AGT ) Media for Bioproduction

Advanced Granulation
Technology
Media
Patented technology enables
complex, chemically-defined media
to be supplied in an innovative,
complete, dry granular format.
• Simplify media preparation by eliminating
the need to add multiple ingredients and adjust
pH and osmolality
• Reduce multiple vendor audits
and qualifications
• Reduce raw material QC testing
and documentation
• Improve inventory management and
warehouse storage efficiency
• Decrease overall process-associated
documentation and training requirements
• Minimize dust generation, reducing clean-up
and possibilities for contamination
• Dissolve instantly for faster mixing time
Media Preparation
Prep with Advanced Granulation Technology Media
AGT Complete Medium
Unit Dose Package
Basal Powder
Medium
Salts II
WFI Quality
Water
NaHCO3
WFI
Water
NaCl for Final
Osmolality Adjustment
COMPLETE CCOMPLETE O M P L E T E M MEDIUM E D I U M
READY-TO-FILTER
RREADY-TO-FILTER E A D Y - T O - F I L T E R
Prep with Dry Powder Media
Growth Factors
e.g. EGF
HCI or NaOH for
pH Adjustment
COMPLETE CCOMPLETE O M P L E T E M MEDIUM E D I U M
READY-TO-FILTER
RREADY-TO-FILTER E A D Y - T O - F I L T E R
In-process
pH Adjustment
Acid
Soluble I
Lipid Supplement
Stock Solution
COMPLETE CCOMPLETE O M P L E T E MEDIUM MMEDIUM E D I U M
READY-TO-FILTER
RREADY-TO-FILTER E A D Y - T O - F I L T E R
NaHCO 3
WFI Quality Water
Addition of NaCl for
Final Osmolality
Adjustment
Protein Supplement
Stock Solution
Insulin and Transferrin
Stock Solutions
Prep with Liquid Media Concentrates
Acid
Soluble II
HCI or NaOH for
final pH Adjustment
Salts I

Sometimes simplification
is the best innovation.
Since the early 1960s, the GIBCO brand has been associated
with breakthrough technologies for cell culture. Several of these
include novel media formats that have been critical to the
evolution of the process: Dry Powder Media (DPM) in 1963,
Liquid Media Concentrates (LMCs) in 1992, and now—in our
40th anniversary year—innovative Advanced Granulation
Technology (AGT) Media.
Invitrogen’s patented AGT process produces a new format of
dry media that can streamline your process from R&D and
Process Development through Production.
AGT media help speed your process
and reduce total cycle cost.
Advanced Granulation Technology produces media in a new,
easy-to-use granular format. The technology enables the
production of dry media for many complex formulations,
providing a complete single-component configuration that is
easy to use and scale up from research through production.
Optimally, for large-scale production, process development
engineers prefer a scalable, serum-free medium in a dry format
requiring minimum supplementation. AGT media can meet
these requirements.
In addition to offering AGT media in a growing variety of
GIBCO catalog formulations, we can also apply our AGT process to your complex custom formulation.
An alternative to liquid and powder.
Compared to other cell culture media formats, the AGT format
offers those involved in process development and manufacturing
many advantages.
AGT media are complete and require no supplementation
or pH adjustment. They eliminate additional procurement,
dispensing, and process steps, and improve set-up in large-scale
media operations.
These media can help reduce total cycle cost. The essential
benefit of a complete AGT medium is that it provides one raw
material for the user, which can decrease costs and time involved
in raw material planning, procurement, and testing. Medium
preparation time is also lessened, because an AGT medium
requires no supplementation and is pH pre-adjusted. Additionally,
because the granules dissolve instantly for faster mixing and
produce less dust, overall medium preparation and clean up
time is reduced.
AGT media offer significant advantages, both technical and
operational, over other cell culture media formats to meet the
needs and requirements of the large-scale production user.
1

Performance-Tested for Growth and Yield
I
2
In cell culture growth and biological production studies of
Hybridomas, CHO, VERO, and HEK 293 cells, GIBCO specialty media derived from the AGT process supported
equivalent performance to identical formulations produced by
traditional liquid and powder processes. (See figures 1 and 2.)
β-Gal Production (units/mL)
IgG Production (ug/mL)
0.4
3.5
0.35
3
0.3
2.5
0.25
0.2
0.15
2
1.5
0.1
1
0.05
0.5
0
0
Day 3 Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 Day 10
Days in Culture
35
30
25
20
15
10
5
0
CD CHO 1X Liquid – β-Gal Production
CD CHO AGT Lot #1 – β-Gal Production
CD CHO AGT Lot #4 – β-Gal Production
CD CHO 1X Liquid – cell growth
CD CHO AGT Lot #1 – cell growth
CD CHO AGT Lot #4 – cell growth
Day 3 Day 4 Day 5 Day 6 Day 7 Day 8
Days in Culture
CD Hybridoma 1X Liquid – IgG Production
CD Hybridoma AGT Lot #1 – IgG Production
CD Hybridoma AGT Lot #2 – IgG Production
Evaluated for Scalability
AGT media are optimally suited to industrial-scale applications.
They demonstrate scalability in characterization studies that
include pH, osmolality, and homogeneity (through HPLC
analysis of amino acids, vitamins, and other components).
(See figures 3 through 9.)
Additionally, data from ongoing real-time stability studies
demonstrate stability of serum-free media made by AGT to
be equivalent to conventional preparation formats.
Performance Scale-up Consistency
CD CHO AGT: Growth and Productivity
Figure 1: Comparable growth and ββ -Gal production
performance is demonstrated with multiple lots of
CD CHO AGT when tested versus 1X Liquid
CD Hybridoma AGT: Growth and Productivity
2.5
2
1.5
1
0.5
Cell growth (viable cells x 10e6/mL)
Cell growth (viable cells x 10e6/mL)
CD Hybridoma 1X Liquid – cell growth
CD Hybridoma AGT Lot #1 – cell growth
CD Hybridoma AGT Lot #2 – cell growth
Figure 2: Comparable growth and IgG production
performance is demonstrated with multiple lots of
CD Hybridoma AGT when tested versus 1X Liquid
0
pH Units
mOsmo
7.6
7.5
7.4
7.3
7.2
7.1
7
6.9
6.8
6.7
6.6
330
325
320
315
310
305
300
9Kg
(n=1)
9Kg
(n=1)
CD CHO AGT: pH
9Kg
(n=1)
9Kg
(n=1)
9Kg
(n=1)
90Kg
(n=6)
90Kg
(n=6)
Lot Size
(n = Number of samples tested)
9Kg
(n=1)
90Kg
(n=6)
90Kg
(n=6)
500Kg
(n=18)
500Kg
(n=18)
500Kg
(n=18)
500Kg
(n=18)
Lot Size
(n = Number of samples tested)
Figure 4: Osmolality within 5 m0smo of target (325)
throughout manufacturing scale-up
330
325
320
315
310
305
300
Figure 3: pH within ±0.1 pH units of target (7.4) throughout
manufacturing scale-up
CD CHO AGT: Osmolality
mOsmo

Percentage of Direct Weigh Control
120
100
140
120
100
80
60
40
20
Percentages of Direct Weigh Control 140
0
Units/mL
80
60
40
20
0
0.2
0.15
0.1
0.05
0
CD CHO AGT: Vitamins
Folic Acid Niacinamide Riboflavin
Mean +/- SEM (n=18)
Thiamine B12
Figure 5: Vitamin concentrations were found to be
within 10% of direct weigh control
Scale-up of CD CHO AGT
Sample #1
Sample #2
Sample #3
Sample #4
Sample #5
Sample #6
Sample #7
Sample #8
Folic Acid
Niacinamide
Riboflavin
Thiamine HCI
L-Histidine
Hydrox-L-Proline
L-Isoleucine
L-Leucine
L-Lysine
L-Methionine
Mean ± SD x 2
(n = Number of samples tested)
3 4 5 6 7 8 9
Days in Culture
9Kg (n=3)
90Kg (n=12)
500Kg (n=36)
Figure 7: Vitamin and amino acid concentrations
were found to be within 10% of direct weigh control
throughout batch scale-up
CD CHO AGT: Productivity
Figure 9: Multiple intra-lot samples demonstrated identical
ββ-Gal productivity
Homogeneity
Percentages of Direct Weigh Control
140
120
100
80
60
40
20
Cells/mL (x10e6)
0
ARG ASP GLM HIS ISO LYS PHE SER TYR TRP
Mean +/- SEM (n=18)
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
CD CHO AGT: Amino Acids
Figure 6: Amino acid concentrations from multiple
intra-lot samples were found to be within 10% of
direct weigh control
CD CHO AGT: Growth
Sample #1
Sample #2
Sample #3
Sample #4
Sample #5
Sample #6
Sample #7
Sample #8
3 4 5 6
Days in Culture
7 8 9
Figure 8: Multiple intra-lot samples demonstrated
identical growth characteristics
3

A New Application for
Fluid Bed Granulation
W
4
We use the pharmaceutical manufacturing technology of fluid
bed granulation to manufacture AGT media. The gentle nature
of fluid bed processing does not affect delicate, biologically
active ingredients. This allows us to do what has never been
done before: manufacture complex, serum-free, protein-free,
and chemically-defined media in a dry format.
In cGMP manufacturing facilities, dry biochemicals are
transferred to a fluid bed processor that suspends the powder
on a column of conditioned air. The suspended powder is
sprayed with a fine mist of aqueous solutions, distributing trace
components homogeneously. As water is evaporated from the
moistened particles, they fuse together into porous granules—a
new form of free-flowing dry media, ready for hydration and
immediate use.
A Novel Granular Format with Unique Properties
Scanning electron micrograph of GIBCO CD CHO AGT granules
The granules produced by the AGT process dissolve instantly and generate
minimal dust. They comprise a free flowing dry medium that is complete and
pH pre-adjusted. Reconstitution with water produces ready to use media,
at target pH and osmolality.
GIBCO Media Available in AGT Format
Chemically-Defined (CD) Media contain no proteins,
hydrolysates, or components of unknown composition. All
components have a known chemical structure, resulting in
consistent product performance and the elimination of
lot-to-lot performance variability.
Protein-Free Media (PFM) are a step closer to defined
formulation as compared to serum-free. A protein-free medium
may still contain undefined components of animal or plant
origin (e.g., various hydrolysates that contribute low molecular
weight peptides).
Serum-Free Media (SFM) do not require supplementation
with serum, but may contain discrete proteins or bulk protein
fractions. In most cases, the protein has been kept to a minimum
and the medium has been optimized for a specific cell type.
To learn how innovative AGT Media
will improve your process, contact your
Invitrogen representative for a detailed
process consultation.
Several GIBCO media formulations, including those listed below,
are available in the AGT format as standard catalog items.
Others are available on a make-to-order basis.
Ordering Information
Description Cat. No. Size
CD CHO AGT 12490-017 1 ✕ 1.L
(Add L-glutamine if required) 12490-025 1 ✕ 10.L
CD Hybridoma AGT 12372-025 1 ✕ 1.L
(Add L-glutamine if required) 12372-017 1 ✕ 10.L
VP-SFM AGT 12559-027 1.✕.1 L
(Add L-glutamine if required) 12559-019 1.✕.10 L
CD 293 AGT 12529-020 1.✕.1 L
(Add L-glutamine if required) 12529-012 1.✕.10 L

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For 40 years, one name has been synonymous
with quality and reliability in cell culture
media, sera, and reagents worldwide.
Printed on recycled paper
These products are for research use, and where appropriate, as raw material
components in further cell culture manufacturing applications. They are not
intended for human or animal diagnostic, therapeutic, or other clinical uses,
unless otherwise stated.
Photograph of fluid bed processor courtesy of Glatt Air Technologies, Inc.,
Ramsey, N.J.; electron micrograph of AGT granules courtesy of McGill
University, Montreal, Quebec.
© 2002 Invitrogen Corporation PAS02-041MS Part No. 332-021646