SlideShare a Scribd company logo
1 of 22
Download to read offline
Optimizing Overall Manufacturing
System Performance through the
Use of Animal-free Cell Culture
Supplements
KiriLynn Svay, Jeff Rosenbloom,
Delyan Rusev, and Matt Croughan

Amgen Bioprocessing Center
Keck Graduate Institute, Claremont, CA

BioManufacturing Summit, January 27, 2010
25 Years of Progress in Clinical Protein Production from Recombinant
                                                                     CHO Cells in Batch and Fedbatch Culture
                                                            Professor Matt Croughan, Keck Graduate Institute, Claremont, CA
                                          100000
                                                                       New to Technology           >2 Years Experience
Product Concentration at Harvest (mg/L)




                                           10000
                                                                          0.205x
                                                             y = 9.941e
                                                               2
                                                              R = 0.7939
                                            1000



                                             100



                                              10
                                                                                                                        0.3477x
                                                                                                           y = 0.082e
                                                                                                              2
                                                                                                            R = 0.9593
                                               1



                                             0.1
                                                   0               5               10          15           20                    25   30
                                                                                        Years Since 1980
Analysis of Variance for Experienced Firms
                                                                Professor M Croughan, Keck Graduate Institute
                                                                           att
                                                  100.0
Percent CV for Product Concentration at Harvest




                                                   90.0

                                                   80.0
                                                                      y = -0.0804x + 50.266
                                                   70.0                      2
                                                                            R = 0.0005
                                                   60.0

                                                   50.0

                                                   40.0

                                                   30.0

                                                   20.0

                                                   10.0

                                                    0.0
                                                          0      5          10           15           20        25   30
                                                                                  Years Since 1980
“..It” flows down hill

                    Consequences of
                      10g/L
   Cell
  Culture
  10g/L
                    • Cell density (volume)
                    • Cell debris
                    • Product Mass
                    • HMW
Purification
                     •Drug
                     Product
                     Development
                                     •From Jon Coffman
Fed-batch cell culture in 1990
Cell culture strategies to optimize
overall system performance
 Consistent and sufficient titers
 ◦ Every process, every time
 Shorter culture duration
 ◦ Higher vol. productivity, lower contamination risk
 Reduced cell death
 ◦ Lower degradative enzyme levels
 ◦ Lower HCP and debris loads downstream
 Improved downstream processing
 ◦ Higher yields or fewer/simpler steps
 Improved product quality and/or stability
Recombinant Human Serum Albumin (rHSA) made in
an animal free production host by InVitria
(www.InVitria.com)
Roles of Albumin in Cell Culture:
◦   Binding and transport mechanism
         Supply of Lipids
         Vitamins
         Hormones
◦   Buffering agent
◦   Detoxifying agent
◦   Protectant from shear
Recombinant Lactoferrin is a naturally occurring iron-
binding protein that was made in an animal free
production system by InVitria (www.InVitria.com)
Advantages of Lactoferrin in Cell Culture
◦ Can be used as a growth factor
◦ Protects against oxidation due to Fe3+ ions
◦ Microbial deterrent
◦ Iron transport to cells
Impact of Cellastim/Lacromin Supplements
  on CHO Cell Culture in CD medium
 % Improvement, Supplemented/Control
  (data from five separate experiments)

 Peak viable cell density (VCD)
 ◦ Average: 28%, Range: 2 - 47%

 Peak titer (product concentration)
 ◦ Average: 40%, Range: 2 – 88%

 Volumetric productivity at peak titer
 ◦ Average: 69%, Range: 3 – 162%
Batch Shake Flask Results
                                                Viable Cell Density
                                 Control                         125:125 mg/L Cellastim:Lacromin
                                 250 mg/L Cellastim              500 mg/L Cellastim
                                                                                                                                   Cells supplemented with 250
                         8                                                                                                         mg/L Cellastim or 500 mg/L
                                                                                                                                   Cellastim reached the greatest
VCD (million cells/mL)




                         7
                         6
                         5
                                                                                                                                   cell density in batch shake
                         4                                                                                                         flasks
                         3
                         2
                         1
                         0
                             0     1       2     3    4   5          6            7              8       9       10       11
                                                              Days
                                                                                                                           % Viability
                                                                                             Control                                       125:125 mg/L Cellastim:Lacromin
                                                                                             250 mg/L Cellastim                            500 mg/L Cellastim
                                                                                       100
                                  This data was very                                    90
                                                                                        80
                                  similar to the InVitria                               70
                                                                         % Viability




                                                                                        60
                                  studies using the same                                50
                                                                                        40
                                  conditions                                            30
                                                                                        20
                                                                                             0       1       2        3        4     5          6   7    8      9    10      11
                                                                                                                                         Days
Bioreactor Density Trends
         Similar trends appeared in the fed batch bioreactors

         •Better growth than un-supplemented cells
         •Slower decline rate at end of culture

                                      Bioreactor Viable Cell Densities and % Viability
                                       125:125 mg/L Cellastim:Lacromin VCD          Control VCD
                                       125:125 mg/L Cellastim:Lacromin %VIA         Control %VIA
                         12                                                                                          100
                                                                                                                     95
                         10
                                                                                                                     90
VCD (million cells/mL)




                                                                                                                     85
                         8
                                                                                                                     80




                                                                                                                           %Viability
                         6                                                                                           75
                                                                                                                     70
                         4
                                                                                                                     65
                                                                                                                     60
                         2
                                                                                                                     55
                         0                                                                                           50
                              0   1        2        3       4        5          6          7       8   9   10   11
                                                                         Days
Specific Glucose and Lactate Trends
                           Specific Glucose Consumption in Fed
                                     Batch Bioreactors                                                                                            In fed batch bioreactors there
                                         Control         125:125 mg/L Cellastim:Lacromin                                                          was decreased sp. glucose
                          0.6                                                                                                                     consumption and decreased
sp. Glucose Consumption




                          0.5                                                                                                                     sp. lactate production from
       (ng/cell*day)




                          0.4
                                                                                                                                                  cells supplemented with
                          0.3
                                                                                                                                                  Cellastim and Lacromin
                          0.2
                          0.1
                           0
                                         0-Prefeed               Post Feed - 7                                               7-10

                                                                                                                                  Specific Lactate Production in Fed
                                                                                                                                           Batch Bioreactors
                                     pH vs.Time
                                                                                                                                      Control      125:125 mg/L Cellastim:Lacromin
                                                                                     sp. Lactate Production (ng/cell*day)


                                    125:125 pH            Control pH
                                                                                                                            1.4
                7.6
                                                                                                                            1.2
                7.4
                7.2                                                                                                          1
pH




                  7                                                                                                         0.8
                6.8                                                                                                         0.6
                6.6
                                                                                                                            0.4
                           0    1    2    3   4      5   6   7   8   9   10 11
                                                                                                                            0.2
                                                     Days
                                                                                                                             0
                                                                                                                                      0-Prefeed           Post Feed - 7              7-10
Glucose Trends EXP4 Bioreactors
                                             Control   250 mg/L Cellastim
                      10.00
                       9.00
                       8.00
Concentration (g/L)




                       7.00
                       6.00
                       5.00
                       4.00
                       3.00
                       2.00
                       1.00
                       0.00
                              0      2   4        6    8         10   12                          14         16       18
                                                           Day

                                                                                                         Lactate Trends EXP4 Bioreactors
                                                                                                                               Control       250 mg/L Cellastim
                                                                                                  4.50
                                                                                                  4.00
                                                                            Concentration (g/L)



                                                                                                  3.50
                                                                                                  3.00
                                                                                                  2.50
                                                                                                  2.00
                                                                                                  1.50
                                                                                                  1.00
                                                                                                  0.50
                                                                                                  0.00
                                                                                                         0        2        4        6    8          10     12     14   16   18
                                                                                                                                              Day
pH Trends EXP4 Bioreactors
                         Control       250 mg/L Cellastim
     7.15
     7.10
     7.05
     7.00
     6.95
pH




     6.90
     6.85
     6.80
     6.75
     6.70
            0    2   4        6    8          10     12         14                  16       18
                                        Day

                                                                                  Osmolality Trends EXP4 Bioreactors
                                                                                                      Control   250 mg/L Cellastim
                                                                                500.00

                                                                                450.00
                                                            Osmolality (mOsm)



                                                                                400.00

                                                                                350.00

                                                                                300.00

                                                                                250.00

                                                                                200.00
                                                                                         0        2   4     6   8         10   12    14   16   18
                                                                                                                    Day
Purification Step Using GE-ÄKTApilot

                                                                         a-IL-8




                                                                         Column
  Column               Sample Loading/                      Eluted
                                             Column Wash                    Re-
Equilibration      Flow-Through Collection                 Fractions
                                                                       equilibration
SDS-PAGE
                               Coomassie Blue Staining
        1        2         3        4        5        6        7        8        9          10
75kD

50kD




25kD




  1    Protein Marker
  2    Supernatant: 125/125 mg/L Cellastim/Lacromin
       Purified a-IL-8 by Protein A Column from supernatant containing 125/125 mg/L
  3
       Cellastim/Lacromin
  4    UF Purified a-IL-8 of #3 product
  5    Supernatant: 250 mg/L Cellastim
  6    Purified a-IL-8 by Protein A Column from supernatant containing 250 mg/L Cellastim
  7    UF Purified a-IL-8 of #6 product
  8    Supernatant: 500 mg/L Cellastim
  9    Purified a-IL-8 by Protein A Column from supernatant containing 500 mg/L Cellastim
 10    UF Purified a-IL-8 of #9 product
SDS-PAGE
                                            Silver Staining
           1        2        3        4        5        6        7        8        9        10

75kD
50kD




25kD




1      Protein Marker
2      Supernatant: 250 mg/L Cellastim
3      Flow – through fraction (time point:10-11min) of #2 run through Protein A Column
4      Purified a-IL-8 by Protein A Column from supernatant containing 250 mg/L Cellastim
5      Waste Fraction (time point: 59-60min) of #2 run through Protein A Column
6      Supernatant from Protein A Column after column disassembling
7      Supernatant: 125/125 mg/L Cellastim/Lacromin
8      Flow – through fraction (time point:10-11min) of # 7 run through Protein A Column
       Purified a-IL-8 by Protein A Column from supernatant containing 125/125 mg/L
9
       Cellastim/Lacromin
10     Waste Fraction (time point: 47-48min) of # 7 run through Protein A Column
ELISA
                                                                    IgG Standard Curve
                                     1200



                                     1000



                                      800




                     Titer (ng/mL)
                                                                      y = 197.13e1.28x
                                                                        R2 = 0.9936
                                      600



                                      400



                                      200



                                        0
                                         0.00       0.20     0.40      0.60       0.80   1.00     1.20   1.40

                                                                         OD 450nm


                                                Amount of IgG loaded                Amount of IgG recovered
     Sample                                                                                                     % Recovered
                                                    (milligrams)                         (milligrams)

   CONTROL
 0 mg/L Cellastim                                          784                                  328                41.8
 0 mg/L Lacromin

125mg/L Cellastim
                                                           619                                  321                51.9
125 mg/L Lacromin


250 mg/L Cellastim                                         537                                  315                58.7

500mg/L Cellastim                                          449                                  313                69.7
Summary
 Optimize overall system performance
 Cellastim/Lacromin supplementation
 ◦   Higher product concentration (titers)
 ◦   Higher specific productivity
 ◦   More efficient glucose metabolism
 ◦   Reduced cell death
 ◦   Higher consistency
 ◦   Equal or improved yields at protein A capture
      Supplements in flow through
      Reduced non-specific losses of MAb

More Related Content

What's hot

Aslo 2012
Aslo 2012Aslo 2012
Aslo 2012emmats
 
120519 mfa acm
120519 mfa acm120519 mfa acm
120519 mfa acmbodewits
 
Results Of Rane Stp Final Report
Results Of Rane Stp Final ReportResults Of Rane Stp Final Report
Results Of Rane Stp Final Reportgueste98444
 
What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...
 What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou... What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...
What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...World Agroforestry (ICRAF)
 
American Pharmaceutical Review Barnes Et Al
American Pharmaceutical Review Barnes Et AlAmerican Pharmaceutical Review Barnes Et Al
American Pharmaceutical Review Barnes Et Albarnes72
 
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...University of the Highlands and Islands
 
Energy scenario and water productivity of maize based CS under CA practices i...
Energy scenario and water productivity of maize based CS under CA practices i...Energy scenario and water productivity of maize based CS under CA practices i...
Energy scenario and water productivity of maize based CS under CA practices i...Joanna Hicks
 
Innovations in Onsite Wastewater Treatment
Innovations in Onsite Wastewater TreatmentInnovations in Onsite Wastewater Treatment
Innovations in Onsite Wastewater TreatmentBuzzards Bay Coalition
 
Deutsche EuroShop | Company Presentation | 07/11
Deutsche EuroShop | Company Presentation | 07/11Deutsche EuroShop | Company Presentation | 07/11
Deutsche EuroShop | Company Presentation | 07/11Deutsche EuroShop AG
 
Deutsche EuroShop | Company Presentation | 09/11
Deutsche EuroShop | Company Presentation | 09/11Deutsche EuroShop | Company Presentation | 09/11
Deutsche EuroShop | Company Presentation | 09/11Deutsche EuroShop AG
 
143. Belmont Stakes
143. Belmont Stakes143. Belmont Stakes
143. Belmont Stakesracingportal
 

What's hot (11)

Aslo 2012
Aslo 2012Aslo 2012
Aslo 2012
 
120519 mfa acm
120519 mfa acm120519 mfa acm
120519 mfa acm
 
Results Of Rane Stp Final Report
Results Of Rane Stp Final ReportResults Of Rane Stp Final Report
Results Of Rane Stp Final Report
 
What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...
 What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou... What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...
What’s cooking on farms? CBD COP 11 - 17 Oct 2012 Domestication of indigenou...
 
American Pharmaceutical Review Barnes Et Al
American Pharmaceutical Review Barnes Et AlAmerican Pharmaceutical Review Barnes Et Al
American Pharmaceutical Review Barnes Et Al
 
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...
Post Medieval land use and vegetation change in upland Ribblesdale, Yorkshire...
 
Energy scenario and water productivity of maize based CS under CA practices i...
Energy scenario and water productivity of maize based CS under CA practices i...Energy scenario and water productivity of maize based CS under CA practices i...
Energy scenario and water productivity of maize based CS under CA practices i...
 
Innovations in Onsite Wastewater Treatment
Innovations in Onsite Wastewater TreatmentInnovations in Onsite Wastewater Treatment
Innovations in Onsite Wastewater Treatment
 
Deutsche EuroShop | Company Presentation | 07/11
Deutsche EuroShop | Company Presentation | 07/11Deutsche EuroShop | Company Presentation | 07/11
Deutsche EuroShop | Company Presentation | 07/11
 
Deutsche EuroShop | Company Presentation | 09/11
Deutsche EuroShop | Company Presentation | 09/11Deutsche EuroShop | Company Presentation | 09/11
Deutsche EuroShop | Company Presentation | 09/11
 
143. Belmont Stakes
143. Belmont Stakes143. Belmont Stakes
143. Belmont Stakes
 

Viewers also liked

Global Spec Overview
Global Spec OverviewGlobal Spec Overview
Global Spec Overviewkbnueml2
 
บทที่ 4 (1)
บทที่ 4 (1)บทที่ 4 (1)
บทที่ 4 (1)einscream
 
Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook
 Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook  Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook
Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook Sara Vega
 
Ελληνικές Επιχειρήσεις και Οικονομική Κρίση
Ελληνικές Επιχειρήσεις και Οικονομική ΚρίσηΕλληνικές Επιχειρήσεις και Οικονομική Κρίση
Ελληνικές Επιχειρήσεις και Οικονομική Κρίσηchaniadevs
 
Reinforcedslab 100917010457-phpapp02
Reinforcedslab 100917010457-phpapp02Reinforcedslab 100917010457-phpapp02
Reinforcedslab 100917010457-phpapp02Laquiao Ganagan
 
Jean baptiste de lamarck (acr)
Jean baptiste de lamarck (acr)Jean baptiste de lamarck (acr)
Jean baptiste de lamarck (acr)aitor_cr
 
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...EMC
 
Texas s ta r powerpoint
Texas  s ta r powerpointTexas  s ta r powerpoint
Texas s ta r powerpointHalogen30
 
профорієнтація і профільне навч.
профорієнтація і профільне навч.профорієнтація і профільне навч.
профорієнтація і профільне навч.Татьяна Глинская
 
Target audience feedback
Target audience feedbackTarget audience feedback
Target audience feedbackharryronchetti
 

Viewers also liked (20)

Global Spec Overview
Global Spec OverviewGlobal Spec Overview
Global Spec Overview
 
บทที่ 4 (1)
บทที่ 4 (1)บทที่ 4 (1)
บทที่ 4 (1)
 
Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook
 Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook  Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook
Venta de Productos Shelo Nabel , @shelonabelshop tienda en facebook
 
Botb14
Botb14Botb14
Botb14
 
Ελληνικές Επιχειρήσεις και Οικονομική Κρίση
Ελληνικές Επιχειρήσεις και Οικονομική ΚρίσηΕλληνικές Επιχειρήσεις και Οικονομική Κρίση
Ελληνικές Επιχειρήσεις και Οικονομική Κρίση
 
Presentation1linx
Presentation1linxPresentation1linx
Presentation1linx
 
Hands on with Hyper-V Clustering Maintenance Mode & Cluster Aware Updating
Hands on with Hyper-V Clustering Maintenance Mode & Cluster Aware UpdatingHands on with Hyper-V Clustering Maintenance Mode & Cluster Aware Updating
Hands on with Hyper-V Clustering Maintenance Mode & Cluster Aware Updating
 
Coworker Crime
Coworker CrimeCoworker Crime
Coworker Crime
 
Reinforcedslab 100917010457-phpapp02
Reinforcedslab 100917010457-phpapp02Reinforcedslab 100917010457-phpapp02
Reinforcedslab 100917010457-phpapp02
 
Euskal Herria
Euskal HerriaEuskal Herria
Euskal Herria
 
Jn wp wcpn2012
Jn wp wcpn2012Jn wp wcpn2012
Jn wp wcpn2012
 
Magne website
Magne websiteMagne website
Magne website
 
Jean baptiste de lamarck (acr)
Jean baptiste de lamarck (acr)Jean baptiste de lamarck (acr)
Jean baptiste de lamarck (acr)
 
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...
RSA Laboratories' Frequently Asked Questions About Today's Cryptography, Vers...
 
летние мастер-классы в сидо
летние мастер-классы в сидолетние мастер-классы в сидо
летние мастер-классы в сидо
 
Texas s ta r powerpoint
Texas  s ta r powerpointTexas  s ta r powerpoint
Texas s ta r powerpoint
 
профорієнтація і профільне навч.
профорієнтація і профільне навч.профорієнтація і профільне навч.
профорієнтація і профільне навч.
 
Modern china
Modern chinaModern china
Modern china
 
Target audience feedback
Target audience feedbackTarget audience feedback
Target audience feedback
 
Cuban revolution
Cuban revolutionCuban revolution
Cuban revolution
 

Similar to Bio manufacturing summit croughan et al jan 2010

Confluence Discovery Technologies
Confluence Discovery TechnologiesConfluence Discovery Technologies
Confluence Discovery TechnologiesCDTBD
 
15th QMOD conference on Quality and Service Sciences 9/07/2012
15th QMOD conference on Quality and Service Sciences 9/07/201215th QMOD conference on Quality and Service Sciences 9/07/2012
15th QMOD conference on Quality and Service Sciences 9/07/2012Brandon Theiss, PE
 
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...CPSA-2012_5-Minutes-Fame
 
22 ionics mass spectrometry - boldly going where no mass spec has gone before...
22 ionics mass spectrometry - boldly going where no mass spec has gone before...22 ionics mass spectrometry - boldly going where no mass spec has gone before...
22 ionics mass spectrometry - boldly going where no mass spec has gone before...CPSA-2012_5-Minutes-Fame
 
Opp for clip 2555 edit
Opp for clip 2555 editOpp for clip 2555 edit
Opp for clip 2555 editnareena4v
 
Mass spec of proteins
Mass spec of proteinsMass spec of proteins
Mass spec of proteinsAmerikAnka
 
SPICE MODEL of UF5401 (Standard Model) in SPICE PARK
SPICE MODEL of UF5401 (Standard Model) in SPICE PARKSPICE MODEL of UF5401 (Standard Model) in SPICE PARK
SPICE MODEL of UF5401 (Standard Model) in SPICE PARKTsuyoshi Horigome
 
Free SPICE Model of SF3L60U in SPICE PARK
Free SPICE Model of SF3L60U in SPICE PARKFree SPICE Model of SF3L60U in SPICE PARK
Free SPICE Model of SF3L60U in SPICE PARKTsuyoshi Horigome
 
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARK
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARKSPICE MODEL of SF3L60U (Standard Model) in SPICE PARK
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARKTsuyoshi Horigome
 
Martin Barden – The audience centre stage | congres podiumkunsten 2012
Martin Barden – The audience centre stage | congres podiumkunsten 2012Martin Barden – The audience centre stage | congres podiumkunsten 2012
Martin Barden – The audience centre stage | congres podiumkunsten 2012Promotie Podiumkunsten
 
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARK
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARKSPICE MODEL of FMLG14S (Professional Model) in SPICE PARK
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARKTsuyoshi Horigome
 
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARK
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARKSPICE MODEL of FMLG14S (Standard Model) in SPICE PARK
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARKTsuyoshi Horigome
 
Wikipedia ws
Wikipedia wsWikipedia ws
Wikipedia wsYu Suzuki
 
Mac Gavery PCSGA_2009
Mac Gavery PCSGA_2009Mac Gavery PCSGA_2009
Mac Gavery PCSGA_2009sr320
 
Multiuser Pre-equalization for Pre-Rake DS-UWB Systems
Multiuser Pre-equalization for Pre-Rake DS-UWB SystemsMultiuser Pre-equalization for Pre-Rake DS-UWB Systems
Multiuser Pre-equalization for Pre-Rake DS-UWB SystemsZahra Ahmadian, PhD
 
An Integer Programming Representation for Data Center Power-Aware Management ...
An Integer Programming Representation for Data Center Power-Aware Management ...An Integer Programming Representation for Data Center Power-Aware Management ...
An Integer Programming Representation for Data Center Power-Aware Management ...Arinto Murdopo
 
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARK
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARKSPICE MODEL of SF3L60U (Professional Model) in SPICE PARK
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARKTsuyoshi Horigome
 
SPICE MODEL of UF5408 (Standard Model) in SPICE PARK
SPICE MODEL of UF5408 (Standard Model) in SPICE PARKSPICE MODEL of UF5408 (Standard Model) in SPICE PARK
SPICE MODEL of UF5408 (Standard Model) in SPICE PARKTsuyoshi Horigome
 

Similar to Bio manufacturing summit croughan et al jan 2010 (20)

Confluence Discovery Technologies
Confluence Discovery TechnologiesConfluence Discovery Technologies
Confluence Discovery Technologies
 
15th QMOD conference on Quality and Service Sciences 9/07/2012
15th QMOD conference on Quality and Service Sciences 9/07/201215th QMOD conference on Quality and Service Sciences 9/07/2012
15th QMOD conference on Quality and Service Sciences 9/07/2012
 
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...
12 2.6 million users can't be wrong keeley murphy and nick duczak - thermo sc...
 
22 ionics mass spectrometry - boldly going where no mass spec has gone before...
22 ionics mass spectrometry - boldly going where no mass spec has gone before...22 ionics mass spectrometry - boldly going where no mass spec has gone before...
22 ionics mass spectrometry - boldly going where no mass spec has gone before...
 
Opp for clip 2555 edit
Opp for clip 2555 editOpp for clip 2555 edit
Opp for clip 2555 edit
 
Starbucks Wait Time Analysis
Starbucks Wait Time AnalysisStarbucks Wait Time Analysis
Starbucks Wait Time Analysis
 
Mass spec of proteins
Mass spec of proteinsMass spec of proteins
Mass spec of proteins
 
SPICE MODEL of UF5401 (Standard Model) in SPICE PARK
SPICE MODEL of UF5401 (Standard Model) in SPICE PARKSPICE MODEL of UF5401 (Standard Model) in SPICE PARK
SPICE MODEL of UF5401 (Standard Model) in SPICE PARK
 
Free SPICE Model of SF3L60U in SPICE PARK
Free SPICE Model of SF3L60U in SPICE PARKFree SPICE Model of SF3L60U in SPICE PARK
Free SPICE Model of SF3L60U in SPICE PARK
 
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARK
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARKSPICE MODEL of SF3L60U (Standard Model) in SPICE PARK
SPICE MODEL of SF3L60U (Standard Model) in SPICE PARK
 
Martin Barden – The audience centre stage | congres podiumkunsten 2012
Martin Barden – The audience centre stage | congres podiumkunsten 2012Martin Barden – The audience centre stage | congres podiumkunsten 2012
Martin Barden – The audience centre stage | congres podiumkunsten 2012
 
237 valeof tiersssp_june07dl
237 valeof tiersssp_june07dl237 valeof tiersssp_june07dl
237 valeof tiersssp_june07dl
 
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARK
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARKSPICE MODEL of FMLG14S (Professional Model) in SPICE PARK
SPICE MODEL of FMLG14S (Professional Model) in SPICE PARK
 
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARK
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARKSPICE MODEL of FMLG14S (Standard Model) in SPICE PARK
SPICE MODEL of FMLG14S (Standard Model) in SPICE PARK
 
Wikipedia ws
Wikipedia wsWikipedia ws
Wikipedia ws
 
Mac Gavery PCSGA_2009
Mac Gavery PCSGA_2009Mac Gavery PCSGA_2009
Mac Gavery PCSGA_2009
 
Multiuser Pre-equalization for Pre-Rake DS-UWB Systems
Multiuser Pre-equalization for Pre-Rake DS-UWB SystemsMultiuser Pre-equalization for Pre-Rake DS-UWB Systems
Multiuser Pre-equalization for Pre-Rake DS-UWB Systems
 
An Integer Programming Representation for Data Center Power-Aware Management ...
An Integer Programming Representation for Data Center Power-Aware Management ...An Integer Programming Representation for Data Center Power-Aware Management ...
An Integer Programming Representation for Data Center Power-Aware Management ...
 
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARK
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARKSPICE MODEL of SF3L60U (Professional Model) in SPICE PARK
SPICE MODEL of SF3L60U (Professional Model) in SPICE PARK
 
SPICE MODEL of UF5408 (Standard Model) in SPICE PARK
SPICE MODEL of UF5408 (Standard Model) in SPICE PARKSPICE MODEL of UF5408 (Standard Model) in SPICE PARK
SPICE MODEL of UF5408 (Standard Model) in SPICE PARK
 

Recently uploaded

DSPy a system for AI to Write Prompts and Do Fine Tuning
DSPy a system for AI to Write Prompts and Do Fine TuningDSPy a system for AI to Write Prompts and Do Fine Tuning
DSPy a system for AI to Write Prompts and Do Fine TuningLars Bell
 
TeamStation AI System Report LATAM IT Salaries 2024
TeamStation AI System Report LATAM IT Salaries 2024TeamStation AI System Report LATAM IT Salaries 2024
TeamStation AI System Report LATAM IT Salaries 2024Lonnie McRorey
 
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptx
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptxMerck Moving Beyond Passwords: FIDO Paris Seminar.pptx
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptxLoriGlavin3
 
Commit 2024 - Secret Management made easy
Commit 2024 - Secret Management made easyCommit 2024 - Secret Management made easy
Commit 2024 - Secret Management made easyAlfredo García Lavilla
 
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024BookNet Canada
 
What's New in Teams Calling, Meetings and Devices March 2024
What's New in Teams Calling, Meetings and Devices March 2024What's New in Teams Calling, Meetings and Devices March 2024
What's New in Teams Calling, Meetings and Devices March 2024Stephanie Beckett
 
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptx
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptxUse of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptx
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptxLoriGlavin3
 
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024BookNet Canada
 
Unleash Your Potential - Namagunga Girls Coding Club
Unleash Your Potential - Namagunga Girls Coding ClubUnleash Your Potential - Namagunga Girls Coding Club
Unleash Your Potential - Namagunga Girls Coding ClubKalema Edgar
 
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptx
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptxPasskey Providers and Enabling Portability: FIDO Paris Seminar.pptx
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptxLoriGlavin3
 
SAP Build Work Zone - Overview L2-L3.pptx
SAP Build Work Zone - Overview L2-L3.pptxSAP Build Work Zone - Overview L2-L3.pptx
SAP Build Work Zone - Overview L2-L3.pptxNavinnSomaal
 
How to write a Business Continuity Plan
How to write a Business Continuity PlanHow to write a Business Continuity Plan
How to write a Business Continuity PlanDatabarracks
 
Dev Dives: Streamline document processing with UiPath Studio Web
Dev Dives: Streamline document processing with UiPath Studio WebDev Dives: Streamline document processing with UiPath Studio Web
Dev Dives: Streamline document processing with UiPath Studio WebUiPathCommunity
 
Nell’iperspazio con Rocket: il Framework Web di Rust!
Nell’iperspazio con Rocket: il Framework Web di Rust!Nell’iperspazio con Rocket: il Framework Web di Rust!
Nell’iperspazio con Rocket: il Framework Web di Rust!Commit University
 
DevEX - reference for building teams, processes, and platforms
DevEX - reference for building teams, processes, and platformsDevEX - reference for building teams, processes, and platforms
DevEX - reference for building teams, processes, and platformsSergiu Bodiu
 
WordPress Websites for Engineers: Elevate Your Brand
WordPress Websites for Engineers: Elevate Your BrandWordPress Websites for Engineers: Elevate Your Brand
WordPress Websites for Engineers: Elevate Your Brandgvaughan
 
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek SchlawackFwdays
 
SALESFORCE EDUCATION CLOUD | FEXLE SERVICES
SALESFORCE EDUCATION CLOUD | FEXLE SERVICESSALESFORCE EDUCATION CLOUD | FEXLE SERVICES
SALESFORCE EDUCATION CLOUD | FEXLE SERVICESmohitsingh558521
 
unit 4 immunoblotting technique complete.pptx
unit 4 immunoblotting technique complete.pptxunit 4 immunoblotting technique complete.pptx
unit 4 immunoblotting technique complete.pptxBkGupta21
 
TrustArc Webinar - How to Build Consumer Trust Through Data Privacy
TrustArc Webinar - How to Build Consumer Trust Through Data PrivacyTrustArc Webinar - How to Build Consumer Trust Through Data Privacy
TrustArc Webinar - How to Build Consumer Trust Through Data PrivacyTrustArc
 

Recently uploaded (20)

DSPy a system for AI to Write Prompts and Do Fine Tuning
DSPy a system for AI to Write Prompts and Do Fine TuningDSPy a system for AI to Write Prompts and Do Fine Tuning
DSPy a system for AI to Write Prompts and Do Fine Tuning
 
TeamStation AI System Report LATAM IT Salaries 2024
TeamStation AI System Report LATAM IT Salaries 2024TeamStation AI System Report LATAM IT Salaries 2024
TeamStation AI System Report LATAM IT Salaries 2024
 
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptx
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptxMerck Moving Beyond Passwords: FIDO Paris Seminar.pptx
Merck Moving Beyond Passwords: FIDO Paris Seminar.pptx
 
Commit 2024 - Secret Management made easy
Commit 2024 - Secret Management made easyCommit 2024 - Secret Management made easy
Commit 2024 - Secret Management made easy
 
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024
Transcript: New from BookNet Canada for 2024: BNC CataList - Tech Forum 2024
 
What's New in Teams Calling, Meetings and Devices March 2024
What's New in Teams Calling, Meetings and Devices March 2024What's New in Teams Calling, Meetings and Devices March 2024
What's New in Teams Calling, Meetings and Devices March 2024
 
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptx
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptxUse of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptx
Use of FIDO in the Payments and Identity Landscape: FIDO Paris Seminar.pptx
 
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024
New from BookNet Canada for 2024: Loan Stars - Tech Forum 2024
 
Unleash Your Potential - Namagunga Girls Coding Club
Unleash Your Potential - Namagunga Girls Coding ClubUnleash Your Potential - Namagunga Girls Coding Club
Unleash Your Potential - Namagunga Girls Coding Club
 
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptx
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptxPasskey Providers and Enabling Portability: FIDO Paris Seminar.pptx
Passkey Providers and Enabling Portability: FIDO Paris Seminar.pptx
 
SAP Build Work Zone - Overview L2-L3.pptx
SAP Build Work Zone - Overview L2-L3.pptxSAP Build Work Zone - Overview L2-L3.pptx
SAP Build Work Zone - Overview L2-L3.pptx
 
How to write a Business Continuity Plan
How to write a Business Continuity PlanHow to write a Business Continuity Plan
How to write a Business Continuity Plan
 
Dev Dives: Streamline document processing with UiPath Studio Web
Dev Dives: Streamline document processing with UiPath Studio WebDev Dives: Streamline document processing with UiPath Studio Web
Dev Dives: Streamline document processing with UiPath Studio Web
 
Nell’iperspazio con Rocket: il Framework Web di Rust!
Nell’iperspazio con Rocket: il Framework Web di Rust!Nell’iperspazio con Rocket: il Framework Web di Rust!
Nell’iperspazio con Rocket: il Framework Web di Rust!
 
DevEX - reference for building teams, processes, and platforms
DevEX - reference for building teams, processes, and platformsDevEX - reference for building teams, processes, and platforms
DevEX - reference for building teams, processes, and platforms
 
WordPress Websites for Engineers: Elevate Your Brand
WordPress Websites for Engineers: Elevate Your BrandWordPress Websites for Engineers: Elevate Your Brand
WordPress Websites for Engineers: Elevate Your Brand
 
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack
"Subclassing and Composition – A Pythonic Tour of Trade-Offs", Hynek Schlawack
 
SALESFORCE EDUCATION CLOUD | FEXLE SERVICES
SALESFORCE EDUCATION CLOUD | FEXLE SERVICESSALESFORCE EDUCATION CLOUD | FEXLE SERVICES
SALESFORCE EDUCATION CLOUD | FEXLE SERVICES
 
unit 4 immunoblotting technique complete.pptx
unit 4 immunoblotting technique complete.pptxunit 4 immunoblotting technique complete.pptx
unit 4 immunoblotting technique complete.pptx
 
TrustArc Webinar - How to Build Consumer Trust Through Data Privacy
TrustArc Webinar - How to Build Consumer Trust Through Data PrivacyTrustArc Webinar - How to Build Consumer Trust Through Data Privacy
TrustArc Webinar - How to Build Consumer Trust Through Data Privacy
 

Bio manufacturing summit croughan et al jan 2010

  • 1. Optimizing Overall Manufacturing System Performance through the Use of Animal-free Cell Culture Supplements KiriLynn Svay, Jeff Rosenbloom, Delyan Rusev, and Matt Croughan Amgen Bioprocessing Center Keck Graduate Institute, Claremont, CA BioManufacturing Summit, January 27, 2010
  • 2. 25 Years of Progress in Clinical Protein Production from Recombinant CHO Cells in Batch and Fedbatch Culture Professor Matt Croughan, Keck Graduate Institute, Claremont, CA 100000 New to Technology >2 Years Experience Product Concentration at Harvest (mg/L) 10000 0.205x y = 9.941e 2 R = 0.7939 1000 100 10 0.3477x y = 0.082e 2 R = 0.9593 1 0.1 0 5 10 15 20 25 30 Years Since 1980
  • 3. Analysis of Variance for Experienced Firms Professor M Croughan, Keck Graduate Institute att 100.0 Percent CV for Product Concentration at Harvest 90.0 80.0 y = -0.0804x + 50.266 70.0 2 R = 0.0005 60.0 50.0 40.0 30.0 20.0 10.0 0.0 0 5 10 15 20 25 30 Years Since 1980
  • 4.
  • 5. “..It” flows down hill Consequences of 10g/L Cell Culture 10g/L • Cell density (volume) • Cell debris • Product Mass • HMW Purification •Drug Product Development •From Jon Coffman
  • 7. Cell culture strategies to optimize overall system performance Consistent and sufficient titers ◦ Every process, every time Shorter culture duration ◦ Higher vol. productivity, lower contamination risk Reduced cell death ◦ Lower degradative enzyme levels ◦ Lower HCP and debris loads downstream Improved downstream processing ◦ Higher yields or fewer/simpler steps Improved product quality and/or stability
  • 8. Recombinant Human Serum Albumin (rHSA) made in an animal free production host by InVitria (www.InVitria.com) Roles of Albumin in Cell Culture: ◦ Binding and transport mechanism Supply of Lipids Vitamins Hormones ◦ Buffering agent ◦ Detoxifying agent ◦ Protectant from shear
  • 9. Recombinant Lactoferrin is a naturally occurring iron- binding protein that was made in an animal free production system by InVitria (www.InVitria.com) Advantages of Lactoferrin in Cell Culture ◦ Can be used as a growth factor ◦ Protects against oxidation due to Fe3+ ions ◦ Microbial deterrent ◦ Iron transport to cells
  • 10.
  • 11. Impact of Cellastim/Lacromin Supplements on CHO Cell Culture in CD medium % Improvement, Supplemented/Control (data from five separate experiments) Peak viable cell density (VCD) ◦ Average: 28%, Range: 2 - 47% Peak titer (product concentration) ◦ Average: 40%, Range: 2 – 88% Volumetric productivity at peak titer ◦ Average: 69%, Range: 3 – 162%
  • 12. Batch Shake Flask Results Viable Cell Density Control 125:125 mg/L Cellastim:Lacromin 250 mg/L Cellastim 500 mg/L Cellastim Cells supplemented with 250 8 mg/L Cellastim or 500 mg/L Cellastim reached the greatest VCD (million cells/mL) 7 6 5 cell density in batch shake 4 flasks 3 2 1 0 0 1 2 3 4 5 6 7 8 9 10 11 Days % Viability Control 125:125 mg/L Cellastim:Lacromin 250 mg/L Cellastim 500 mg/L Cellastim 100 This data was very 90 80 similar to the InVitria 70 % Viability 60 studies using the same 50 40 conditions 30 20 0 1 2 3 4 5 6 7 8 9 10 11 Days
  • 13. Bioreactor Density Trends Similar trends appeared in the fed batch bioreactors •Better growth than un-supplemented cells •Slower decline rate at end of culture Bioreactor Viable Cell Densities and % Viability 125:125 mg/L Cellastim:Lacromin VCD Control VCD 125:125 mg/L Cellastim:Lacromin %VIA Control %VIA 12 100 95 10 90 VCD (million cells/mL) 85 8 80 %Viability 6 75 70 4 65 60 2 55 0 50 0 1 2 3 4 5 6 7 8 9 10 11 Days
  • 14. Specific Glucose and Lactate Trends Specific Glucose Consumption in Fed Batch Bioreactors In fed batch bioreactors there Control 125:125 mg/L Cellastim:Lacromin was decreased sp. glucose 0.6 consumption and decreased sp. Glucose Consumption 0.5 sp. lactate production from (ng/cell*day) 0.4 cells supplemented with 0.3 Cellastim and Lacromin 0.2 0.1 0 0-Prefeed Post Feed - 7 7-10 Specific Lactate Production in Fed Batch Bioreactors pH vs.Time Control 125:125 mg/L Cellastim:Lacromin sp. Lactate Production (ng/cell*day) 125:125 pH Control pH 1.4 7.6 1.2 7.4 7.2 1 pH 7 0.8 6.8 0.6 6.6 0.4 0 1 2 3 4 5 6 7 8 9 10 11 0.2 Days 0 0-Prefeed Post Feed - 7 7-10
  • 15. Glucose Trends EXP4 Bioreactors Control 250 mg/L Cellastim 10.00 9.00 8.00 Concentration (g/L) 7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00 0 2 4 6 8 10 12 14 16 18 Day Lactate Trends EXP4 Bioreactors Control 250 mg/L Cellastim 4.50 4.00 Concentration (g/L) 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00 0 2 4 6 8 10 12 14 16 18 Day
  • 16. pH Trends EXP4 Bioreactors Control 250 mg/L Cellastim 7.15 7.10 7.05 7.00 6.95 pH 6.90 6.85 6.80 6.75 6.70 0 2 4 6 8 10 12 14 16 18 Day Osmolality Trends EXP4 Bioreactors Control 250 mg/L Cellastim 500.00 450.00 Osmolality (mOsm) 400.00 350.00 300.00 250.00 200.00 0 2 4 6 8 10 12 14 16 18 Day
  • 17.
  • 18. Purification Step Using GE-ÄKTApilot a-IL-8 Column Column Sample Loading/ Eluted Column Wash Re- Equilibration Flow-Through Collection Fractions equilibration
  • 19. SDS-PAGE Coomassie Blue Staining 1 2 3 4 5 6 7 8 9 10 75kD 50kD 25kD 1 Protein Marker 2 Supernatant: 125/125 mg/L Cellastim/Lacromin Purified a-IL-8 by Protein A Column from supernatant containing 125/125 mg/L 3 Cellastim/Lacromin 4 UF Purified a-IL-8 of #3 product 5 Supernatant: 250 mg/L Cellastim 6 Purified a-IL-8 by Protein A Column from supernatant containing 250 mg/L Cellastim 7 UF Purified a-IL-8 of #6 product 8 Supernatant: 500 mg/L Cellastim 9 Purified a-IL-8 by Protein A Column from supernatant containing 500 mg/L Cellastim 10 UF Purified a-IL-8 of #9 product
  • 20. SDS-PAGE Silver Staining 1 2 3 4 5 6 7 8 9 10 75kD 50kD 25kD 1 Protein Marker 2 Supernatant: 250 mg/L Cellastim 3 Flow – through fraction (time point:10-11min) of #2 run through Protein A Column 4 Purified a-IL-8 by Protein A Column from supernatant containing 250 mg/L Cellastim 5 Waste Fraction (time point: 59-60min) of #2 run through Protein A Column 6 Supernatant from Protein A Column after column disassembling 7 Supernatant: 125/125 mg/L Cellastim/Lacromin 8 Flow – through fraction (time point:10-11min) of # 7 run through Protein A Column Purified a-IL-8 by Protein A Column from supernatant containing 125/125 mg/L 9 Cellastim/Lacromin 10 Waste Fraction (time point: 47-48min) of # 7 run through Protein A Column
  • 21. ELISA IgG Standard Curve 1200 1000 800 Titer (ng/mL) y = 197.13e1.28x R2 = 0.9936 600 400 200 0 0.00 0.20 0.40 0.60 0.80 1.00 1.20 1.40 OD 450nm Amount of IgG loaded Amount of IgG recovered Sample % Recovered (milligrams) (milligrams) CONTROL 0 mg/L Cellastim 784 328 41.8 0 mg/L Lacromin 125mg/L Cellastim 619 321 51.9 125 mg/L Lacromin 250 mg/L Cellastim 537 315 58.7 500mg/L Cellastim 449 313 69.7
  • 22. Summary Optimize overall system performance Cellastim/Lacromin supplementation ◦ Higher product concentration (titers) ◦ Higher specific productivity ◦ More efficient glucose metabolism ◦ Reduced cell death ◦ Higher consistency ◦ Equal or improved yields at protein A capture Supplements in flow through Reduced non-specific losses of MAb