Purification Department

Suspension
• Albumin: Around (850 to 900kg).
• CWFI (Water for Injection) same
quantity of albumin. (first 70%
and 30%)
• Mixing time: 60 min.
• pH adjustment with NaOH
• pH= 6.9 to 7.1
• IR: 16% protein

•Note: 58.8 Kg of NaOH solution.

•Normally 3 additions but the
second one and the third one are
10kg and 5 kg respectively.
Clarification
• CUNO Filter preparation (Flushing
2x50 LA=> 30 min HWFI and 30 min
CWFI; Flushing 1x90 LA => 15 min
CWFI and 15 min HWFI).
• Conductivity after the flushing
process ( Value: < 3uS/cm)
• Filter Aid addition before starting
clarification.
• Turbidity value during the
clarification must be less than 9
NTU.
• CWFI Addition to obtain 10% of
protein.
• 300kg in V1.1 and the remaining
water in V1.2A.
Diafiltration
and Heat Shock
• UF-120 preparation (CIP
and Rinses).
• Before UF we must run:
• NWP: Normalized
Water Permeability.
• TDF: Diffusion Test or
Integrity Test.
• LAL sample (<240
IU/mL)
• Connection of UF-120 to
Vessel 1.2A.
Diafiltration and Heat Shock

• UF-120 filtration.
• IR value before starting is more and less
10%
• IR value to achieve is 14%
• Concentration Phase.
Diafiltration
and Heat Shock
• 2.5N NaCl required
• Total volume discard during
diafiltration.
• NaCl buffer solution (440 to
490 mOsm/.Kg)
Diafiltration
and Heat Shock
• Post- diafiltration
Concentration
• IR will be around 15% to 18%
after diafiltration.
• Final Volume is 14%
• Post wash: addition CFWI
and clean all the flow
connection process related
with UF.
Diafiltration and Heat Shock
• Stabiliser Calculation:
• Sodium Caprylate
• N-acetyl Tryptophan
• 2N NaOH solution
• Stabiliser Solution pH:
7.0 to 7.5
• Addition 1L/1min.
Heat Shock
• Temperature: 59.5 to 60.5
degrees.
• Start the Eurotherm
Electronic Recorded.
• Time: 3hr with 30minutes.
• Process starts when we get
59.5 degrees in PCS7.
Second
Clarification
• Flushing CUNO preparation
2x90 LA (CWFI x 30 min and
HWFI x 30 min.)
• Conductivity after the
flushing process ( Value: <
3uS/cm).
• Turbidity value during the
clarification must be less
than 8 NTU.
• Post flush volume taken
from PCS7.
 Formulation 5%
• Process to obtain the specific
percentage of protein.
• IR sample will be around 12 to
14%.
• Calculate the CWFI needed to
get 5.17%
• Add 80% of that volume of
water.
• IR sample to check % of
protein.
• Calculate final moles NaCl
needed for 5.17%
• IR measurement to check %
of protein.
• Determinate the volume of
CWFI to achieve 5.05% of
protein.
• Conductivity (< 2uS/cm)
• pH Solution ( 7.05 to 7.15)
• Osmolality (210-240
mOsm/Kg).
 Vent Filter Preparation. (Vent Filter Test)

Sterile
Sterilizing Filter Preparation. ( Flushing time and Sterilizing
Filter Test)
Autoclave Cycle (Equipment necessary to the set up)

Filtratio Pressure Gauge ( Validated)

CIP (Clean in Place) including Manual Clean (J-tube) and

n Getinge Machine Cycle Number.

SIP (Steam in Place)

Standardization Scales.

Pre-filter Preparation.
Sterile Filtration

Scale 5 Scale 6