CDM4PERMAb
HYCLONE MEDIA AND SUPPLEMENTS

HyClone™ CDM4PERMAb medium has a chemically definedand animal-derived component-free (ADCF) formulation.
This high-performance cell culture medium has been developed
through the Metabolic Pathway Design process (see box) toincrease process yields in the production of human antibodies
and recombinant proteins using PER.C6™ technology. The medium
has been successfully tested in a variety of applications, including
fed-batch bioreactor cultures. CDM4PERMAb is available in liquidand powder formats in user-friendly packaging (Fig 1).
Key features of CDM4PERMAb include•

Animal-derived component-free•

Chemically defined formulation•

Designed for high cell yield and recombinant protein
production•

Allows for direct or sequential adaptation•

Component traceability•

Manufactured according to cGMP guidelines

Fig 1. CDM4PERMAb medium is available as liquid or powder in packsizes suitable for small-volume cell culture as well as large-scale
bioprocessing applications.

Specifications•Does not contain L-glutamine

Metabolic Pathway Design process•

Does not contain phenol red•

Liquid medium contains poloxamer 188•

Store medium at 2°C to 8°C, away from light•

Powder medium should be stored protected from moisture
in a tightly sealed container

An optimal cell culture process is dependent of a variety offactors including the parental cell line, the genetic makeupof the specific clone, medium and feed composition, as
well as process variables to maximize viable cell densities
and titers while maintaining cell morphology. Our experts
in medium design and development know and understandhow these factors can influence the metabolic processes
involved. They evaluate the culture’s metabolic activities,
measuring nutritional demand and waste creation to makesure the correct type and quantity of nutrients are used
to minimize waste and resultant cell toxicity. Our experts
use their understanding of metabolic pathways to optimize
medium composition for enhanced productivity and viable
cell densities. Once a medium has been optimized using
this Metabolic Pathway Design process, our scientists canhelp you devise the most effective cell culture strategy
using a combination of medium and feeds to further enrichproductivity and reduce process inefficiencies.

Suggested preparation
Reconstitution of CDM4PERMAb powder medium
1. While stirring, add CDM4PERMAb powder medium to cell
culture-grade water at 90% of final preparation volume(17.3 g/L). If your water source is normally cool, it may be
useful to adjust the water temperature. Using warmer room
temperature water (22°C to 25°C) will improve solubilization
time. Mix for 20 min or until dissolved. Medium should be aclear, golden solution at this point.

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2. Add 0.5 g/L of poloxamer 188 and 3.2 g/L of sodium
bicarbonate. Ensure each component has completely
dissolved before adding the next component.
3. Bring vessel to final volume with culture grade water.
Allow solution to mix for 10 to 20 min.
4. Adjust pH to between 7.0 and 7.2 by adding 1 N NaOH,
or 1 N HCl drop wise to solution.
5. Check osmolality. Expected value is 290 to 340 mOsm/kg.
6. Sterile filter into desired container using a 0.2 µm sterile filter.

Preparation notes
CDM4PERMAb powder medium does not contain L-glutamine.
Recommended concentration: 4 mM.

General culture recommendations1. Cultures should be incubated at 37°C in a 5% CO2 environment.
2. Maintain stock cells in classical medium with serum
supplementation.

Quality control testing
Quality control test specifications are listed in Table 1.
Table 1. Test specifications1

Appearance
Osmolality
pH
Sterility
Endotoxin
Application
1

Clear yellow solution
290 to 340 mOsm/kg
7.0 to 7.4
No growth (bacteria or fungi)≤ 1.0 EU/mL1
Growth promotion

Refer to certificate of analysis for actual results.

Custom production
Formulations and delivery systems can be customized toyour specific process requirements or optimized to maximize
process yields.

Rapid Response Production (RRP)

3. Upon subsequent passage, following trypsinization, seed
two new T-flasks. One flask will be maintained at the current
serum concentration as a backup, while the other will
be reduced to half that serum concentration. This serum
depletion process is repeated until reaching a concentrationof 2.5% fetal bovine serum (FBS).

Our RRP program manufactures up to 200 L of your custom
prototype formulation within seven working days of your request.
Use our RRP service to expedite the development and testing ofcustom buffers and process liquids for your biopharmaceuticalmanufacturing process.

4. Re-plate cells at 2.5% FBS and incubate for 24 h to allow
attachment and spreading.

Related products

5. Following the 24 h incubation, replace the serum-containing
medium with CDM4PERMAb medium. Cells will not attach
firmly and should be passaged without trypsin.
6. A passage schedule of 3 to 4 days should be maintained for 2 to3 passages. Adaptation is complete once cells have transitioned
to a doubling rate of 24 h or less per doubling.

Cell maintenance
Maintain adapted cells by establishing a passage schedulethat allows the cells to be passed while in log growth phase.
PER.C6 cells cultivated in CDM4PERMAb medium for routine
maintenance should be subcultured every 72 to 96 h.
The passage schedule and seeding density may be adjusted
to optimize performance. The recommended population
seeding density of new cultures for general maintenance is
0.3 × 106 cells/mL. The culture viability of an adapted culture
should remain greater than 90%. During adaptation fromserum-containing medium, however, viabilities might be slightly
lower than 90%. Cells should exhibit a population doubling timeof approximately 24 to 30 h. If the recommended population seeddensity of 0.3 × 106 cells/mL is used, cultures typically reach peakcell population densities of between 8.0 and 12.0 × 106 cells/mL,depending on the specific clone used. Doubling times during anadaptation period might be higher.

Cryopreservation
CDM4PERMAb medium adapted cells can be cryopreservedin a medium consisting of a 1:1 ratio of fresh and conditioned
CDM4PERMAb medium. To this, add DMSO to a final
concentration of 7.5%.

2

Table 2 gives an overview of HyClone supplements.

HyClone Cell Boost kit
Cell Boost™ Process Supplements (100 g each) contain samples
of supplements designed to increase cell productivity in a varietyof cell lines (Table 2). Each supplement is developed through theMetabolic Pathway Design process and is chemically defined andprotein-free with no animal-derived components.

HyClone LS250 supplement
LS250 is a chemically defined, animal-derived component-free lipid
supplement developed to stimulate cell growth and monoclonal
antibody (MAb) production in NS0 cell cultures using traditional
hybridoma serum-free media.

HyClone LS1000 supplement
LS1000 supplement is a chemically defined, animal-derived
component-free lipid supplement developed to stimulate cell
growth and MAb production in NS0 cell cultures using traditional
hybridoma serum-free media.
The supplement is formulated using a proprietary complexing
process for enhanced cholesterol delivery. LS1000 has beensuccessfully tested in a variety of serum-free medium cultures,
including HyClone CDM4NS0 and CDM4MAb media.

Table 2. Supplement matrix
Aminoacids
Cell Boost 1
Supplement (R05.2)
Cell Boost 2
Supplement (R15.4)
Cell Boost 3
Supplement (JM3.5)
Cell Boost 4
Supplement (PS307)

VitaminsGlucose•

•
•
•
•

•
•
•
•

•
•

Traceelements

Growthfactors••

Hypoxanthine/thymidine

ADCF*
lipids

ADCF*
cholesterol

•
•

•

•

Cell Boost 5
Supplement (CN-F)

••

•

•

•

•

•

•

Cell Boost 6
Supplement (CN-T)

••

•

•

•

•

•

•

•

•
•

LS250 supplement
LS1000 supplement

Suitablefor
HEK293CHO
PER.C6
CHO
Hybridoma
Myeloma
CHO

CodenumberSH30584
SH30596
SH30825
SH30857

Hybridoma
NS0HEK293
CHOT-Cells
Hybridoma
NS0HEK293
CHO

SH30865

SH30866

NS0SH30554

NS0SH30555* Animal-derived component-free

Ordering information
CDM4PERMAb is manufactured in homogenous liquid lot sizes up
to 10 000 L and powder lots up to 250 000 L.
ProductSize

HyClone CDM4PERMAbliquid medium

500 mL bottle
1000 mL bottle
10 L bag
20 L bag
50 L bag
100 L bag
200 L bag
500 L bag
900 L bag

SH30871.01
SH30871.02
SH30871.05
SH30871.06
SH30871.07
SH30871.08
SH30871.09
SH30871.10
SH30871.11

HyClone CDM4PERMAb
powder medium

1 × 5 L*1 × 10 L*
1 × 50 L*
1 × 100 L*
1 × 500 L†
1 × 1000 L†

SH30872.01
SH30872.02
SH30872.03
SH30872.04
SH30872.05
SH30872.06* High-density polyethylene (HDPE) bottle†
Polybag/pailcytiva.com/hyclone
Cytiva and the Drop logo are trademarks of Global Life Sciences IP Holdco LLC
or an affiliate. Cell Boost and HyClone are trademarks of Global Life Sciences
Solutions USA LLC or an affiliate doing business as Cytiva.
PER.C6 is a trademark of Crucell. All other third party trademarks are theproperty of their respective owners.
© 2020 Cytiva
All goods and services are sold subject to the terms and conditions of saleof the supplying company operating within the Cytiva business. A copy of
those terms and conditions is available on request. Contact your local Cytiva
representative for the most current information.
For local office contact information, visit cytiva.com/contact

CY12696-10Jul20-DF

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Size

HyClone Cell Boost kit

6 × 100 g

HyClone LS1000
cholesterol supplement

50 mL bottle

SH30554.01

100 mL bottle
500 mL bottle
1000 mL bottle

SH30554.02
SH30554.03
SH30554.04

HyClone LS250
lipid supplement

100 mL bottle
500 mL bottle
1000 mL bottle

SH30555.01
SH30555.02
SH30555.03

Code number

Code number
SH30890