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LONG®R3IGF-I
Manufactured by Repligen Distributed exclusively by SAFC
GMP Growth Factor Additive for Cell Culture Manufacturing
sigma-aldrich.com/safc
Maximize Productivity in your Chemically-Defined Cell Culture Media
Growth Factor Additive for Cell Culture ManufacturingLONG®R3IGF-I: Meeting Market Needs
Increase Cell DensityGreater Cell Productivity
Maintain Higher Viability
Industry Need SAFC Feature Benefit
Greater Cell Productivity
Increased cell density Promotes cell proliferation and higher titer
Maximizes growth in cell culture applications
Maintain higher viability Greater cell culture stability
Prolongs cell culture activity
Optimize Cost in Use
Highly potent High titer and lower cost in use
200x more effective than insulin
Reduced risk with greater performance
Regulatory Compliant
Manufactured according to applicable GMP guidelines (ICH Q7)
Designed for use in cell culture manufacturing
Meets highest regulatory criteria for raw materials
Trusted supply chain 20 year track record of high quality and consistency
Validated and controlled manufacturing processes
Reduced risk by management of change
Animal-free Ideal for animal-free, chemically-defined products
Actively used in regulatory–approved commercial products
Regulatory piece of mind
Established Supply Chain
Readily available inventory Security of supply: Available for immediate shipment
Cold chain storage Kept in controlled cold storage until ready to ship
Animal-Free
Prod
uctiv
ity
Serum (FBS)Optimum cell growth
& productivity
Serum-freeProtein-richGood cell growth
& productivity
Protein-freeChemically-de�ned
Sub-optimal cell growth& productivity
RepligenCell Culture
Recombinant Animal-free Proteins
It is often hard to maintain or improve productivity in serum or animal-free formulations. Many biopharmaceutical manufacturers are trying to eliminate animal-containing raw materials, or move
Improve Productivity and Cell Culture Stability
Use the Growth Factor Designed for Commercial Cell CultureGrowth factors are essential for long-term growth and proliferation of cell lines in serum-free media formulations. A common growth factor for CHO cells and stem cell culture is insulin. Although insulin is used as a supplement, its primary use is as a therapeutic for the treatment of diabetes. This has led to supply and availability issues for cell culture users. In contrast, LONG®R3IGF-I is a dedicated raw material manufactured exclusively for the cell culture market and provides a consistent, GMP compliant, and reliable growth factor. It is produced in an Escherichia coli expression system without the use of animal-derived components.
LONG®R3IGF-I optimizes growth in animal-free cell cultures
towards chemically-defined media to reduce risk of contamination. LONG®R3IGF-I is a highly potent recombinant animal-free material that helps reduce risk while improving productivity.
Proven Science to Help Maximize your Cell Culture PerformanceLONG®R3IGF-I, an analogue of human insulin-like growth factor I (IGF-I), is a highly potent & effective animal-free recombinant growth factor specifically engineered for use in industrial cell culture. Insulin and IGF-I have a similar amino acid sequence and protein structure, allowing both to bind to each other’s receptor with relatively low affinity.
It is widely accepted that in CHO cells the effects of insulin are mediated by the IGF-IR, due to the fact there are relatively few IR present on CHO cells and that insulin must be present at a high, non-physiological concentration, typically 1–10 mg/L.
A more effective growth factor is one which targets and activates the IGF-IR directly, such as native IGF-I or its analogue, LONG®R3IGF-I. The substitution of an arginine for glutamic acid at position in conjunction with the 13 amino acid N-terminal extension peptide, provides LONG®R3IGF-I a distinct biological advantage resulting in > 1000-fold reduced affinity for IGFBPs enhancing bioavailability and effectiveness in comparison to native IGF-I.
Fig. 3
200%
0%
50%
100%
0 7
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150%
9 14
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elat
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ay-1
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Cont
rol)
Prolonged Cell Culture Activity Through Greater StabilityUnder normal cell culture conditions, LONG®R3IGF-I is more stable than insulin, persisting up to two times longer. Insulin is degraded in cell culture by enzymes (insulinases) secreted by cells into culture media. In addition, insulin is more rapidly internalized and degraded compared to IGF-1 and LONG®R3IGF-I. The extended cell culture stability of LONG®R3IGF-I results in prolonged activity and associated benefits to cell culture (Figure 3).
Gain Greater Stability and Enhanced PerformanceIncrease Cell Density and Titer, Maintain Higher Viability and Extend Culture Duration
Stress-induced apoptosis is the major cause of loss of cell viability. Activation of the IGF-IR results in the stimulation of a number of signal transduction cascades that have been identified as important for cell survival and proliferation.
Increased Cell Density and Titer using LONG®R3IGF-I Growth Factor
LONG®R3IGF-I Provides Higher Titer than Insulin
Use of LONG®R3IGF-I Results in Prolonged Cell Culture Duration vs. Insulin
100
0
20
40
60
0 2 4 6 8 10
Time (Days) in Culture
80
= Long® R3 IGF-I= Insulin
12
120
% R
ecov
ery
Fig. 1
In order to confirm that the increases in cell growth and titer seen in spin tubes translated to bioreactors, two reactors were run side-by-side. One contained DP-12 cells adapted to chemically-defined media without growth factor in unsupplemented chemically-defined media, and the other contained DP-12 cells adapted to LONG®R3IGF-I in chemically-defined media with 100 ng/mL growth factor. Reactors were run as described in the Methods section. LONG®R3IGF-I supplementation resulted in a 1.4-fold increase in growth and a 2.2-fold increase in volumetric productivity versus the unsupplemented reactor, Figure 1 and 2.
Fig. 4
Fig. 2
LONG®R3IGF-I not only results in greater activation of the IGF-IR over insulin, but also results in greater activation of key antiapoptotic and proliferative signaling molecules; Akt and MAPK. Increased activation of these signaling molecules prevents the loss of viability caused by different culture conditions.
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= 100 ng/mL LR3= No Growth factor
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Days in Culture
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= 100 ng/mL LR3= No Growth factor
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250%
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150%
0 5
Days in Culture
200%
= 100 ng/mL LR3= No Growth factor
10 15 20
Perc
etag
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-17
no G
F Co
ntro
l)250%
0%
50%
100%
150%
0 5
Days in Culture
200%
= 100 ng/mL LR3= No Growth factor
10 15 20
Perc
etag
e Ti
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-17
no G
F Co
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Figure 3: LONG®R3IGF-I or insulin levels in CHO culture over time. Levels were measured by ELISA and normalized to initial concentration, since LONG®R3IGF-I is added at ng/mL concentrations and insulin at µg/mL levels. LONG®R3IGF-I levels could be measured in culture via ELISA after 11 days, while insulin was rapidly cleaved by secreted proteases and not measurable after only 6 days.
Figure 4: Titer of DP-12 cells grown in chemically-defined media using spin tubes. Bars represent triplicate cultures. A 62% increase in titer was seen for 100 ng/mL supplementation vs no GF.
Regulatory Compliant, GMP and Animal-FreeLONG®R3IGF-I is manufactured under GMP in compliance with the International Conference on Harmonisation (ICH) Q7A guidelines. The manufacturing plant is regularly audited by major European, US and Japanese biopharmaceutical companies. LONG®R3IGF-I is produced in a proprietary E.coli fermentation process that is completely animal-free. No animal derived materials are used during the manufacture or storage of LONG®R3IGF-I. A certificate of origin is available on request.
A unique peptide, LONG®R3IGF-I has been manufactured and supplied to the cell culture market for over 20 years. In addition, LONG®R3IGF-I is used in the manufacture of several FDA, EMA, and MHW approved marketed products, with many more in late-phase clinical trials.
Security of Supply Backed by a Comprehensive Risk Management StrategyLONG®R3IGF-I inventory is built in advance to handle the fluctuating needs of customers and is available for immediate shipment. A comprehensive risk management strategy includes an inventory surplus equal to 2 year’s forecast demand and is maintained in secured storage facilities in the U.S. and Europe. For a complete overview of the risk management strategy, contact your SAFC sales representative.
Broad Use Across Cell LinesLONG®R3IGF-I has been used with numerous cell types including CHO, vaccine-producing cell lines, fibroblasts and stem cells. All cell types that have a growth response to insulin in cell culture have the potential to respond to LONG®R3IGF-I. LONG®R3IGF-I provides equivalent or better performance to recombinant insulin depending on the cell line and clone.
User-Friendly Formulations for Greater FlexibilityAvailable as either a lyophilized powder or a liquid formulation, both formats are stable at 2-8º C. The liquid formulation is ready to use with no need to defrost or reconstitute. Simply open and dilute directly into cell culture media. LONG®R3IGF-I can also be milled into media available from SAFC.
Products:GMP quality (media grade)*
Product Number
Pack sizes-SKU
Powder 85580C 85580C-5mg
85580C-50mg
Solution (1 mg/mL in 0.1 M acetic acid)
91590C 91590C-5mL 91590C-100mL
Store at 2-8° C
* All products made under GMP guidlines, but only larger pack sizes have full GMP validation for use in manufacturing.
How to Order: Visit sigmaaldrich.com and enter the selected product number in search bar, or contact your SAFC sales representative.
Designed for Commercial Manufacturing
SAFC International SitesGlobal Email: [email protected]
sigma-aldrich.com/safc
KLQ822801074
©2014 Sigma-Aldrich Co. LLC. All rights reserved. SAFC and SIGMA-ALDRICH are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. LONG is a registered trademark of Repligen Sweden AB. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and
conditions may apply. Please see product information on the SAFC website at www.safcglobal.com and/or on the reverse side of the invoice or packing slip.
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