This document discusses Lonza's efforts to improve cleaning of buffer preparation equipment rings caused by slip agents used in plastic bag liners. It summarizes: 1) Lonza initially validated use of ProKlenz Booster peroxide cleaner, which successfully removed slip agents but caused some surface oxidation. Occasional residual rings required manual cleaning. 2) Further testing identified a non-ionic surfactant cleaner that outperformed ProKlenz Booster at removing a surrogate residue on coupons. 3) Next steps include generating a more accurate residue for additional coupon tests, scaling the improved cleaner for at-scale buffer preparation vessel trials, and continuing efforts with suppliers to reduce or eliminate slip agent use.

1. Pharma&Biotech
Andrew Harris / Lonza Biologics Inc., Portsmouth, NH 03801 / 16 September 2014©
Lonza
CIP of Buffer Preparation “Rings”
Annual CIP Summit 2014
Pharma&Biotech

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Sep-14
Outline
 Lonza
 What are Buffer Preparation “Rings” ?
 Supply Chain
 Impact
 ClP
 Buffer Ring CIP version 1.0
 Buffer Ring CIP version 2.0
 Next Steps

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Sep-14
Lonza
 Lonza is a leading custom manufacturer of active
pharmaceutical ingredients, intermediates and biotechnology
solutions on the basis of advanced chemical synthesis, microbial
fermentation and mammalian cell culture, which it supplies to the
life sciences industry.
 Headquartered in Basel,
Switzerland.
 40 major production and
R&D Facilities worldwide
employing approximately
10,000.

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Sep-14
Lonza Biologics - Portsmouth
 Located at the Pease International Tradeport in Portsmouth, NH
 Contract Manufacturing Organization
 Custom manufacturer of therapeutic monoclonal antibodies and
recombinant proteins using mammalian cell culture
 Licensed multi-product facility
 Clinical and Commercial
 5k and 20k capacity
 700+ employees - 24/7 operation
 cGMP Manufacturing since 1996

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Sep-14
What are Buffer Prep “Rings?”
 Form only at the Air-Liquid Interface (ALI), on the order of 0.5-3cm thick
after a little as one buffer makeup
 Slip Agents
 Used in bag manufacturing to reduce friction between film and equipment +
adjacent film
 Fatty acid amides: Most important slip agents for polyolefins (PE, PP, etc)
 The raw materials are not ‘pure’ chemicals so have a molecular weight range
 Anti-Block Agents
 Used in bag manufacturing to prevent wraps of a roll of film from sticking to each
other
 Inorganic – Talc (Magnesium Silicate), Silicon Dioxide, Calcium Carbonate
 Polymers
 Likely impurities of buffer raw materials / salts
 Polyolefins (PE, PP), Fluoropolymers (PTFE), etc
 Residual buffer materials / salts

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Sep-14

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Sep-14
Slip Agents
 Hydrocarbons of various saturation with amide group. Most are
mono-unsaturated
 Slip agents’ raw materials generally contain some amount of
various other slip agents
 Difficult to identify specific slip agents on FTIR
NH2
O
NH2
O
Erucamide (13-cis-docosenamide)
Oleamide (9-cis-octadecenamide)

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Sep-14
Anti-Block agents
 Combat friction and Van der Waals forces
during film blowing & casting
 Added directly to polymer resin, migrates to
surface during film manufacturing. Anti-block
components melt at much higher temperatures
than the polymers
 Slip-agent-free plastics still contain
Anti-Block agents
 Upon reducing Slip Agent content,
additional Anti-Block is added

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Sep-14
Supply Chain
 Lonza has contacted raw material suppliers (chemicals and
bags) and identify which use slip agents in their bag liners
 Questionnaire to suppliers
 Slip agent usage information 2 levels back in supply chain
 Request to work with us to remove slip agents from their process
 Few stated no, citing L/E data
 Some who stated yes, later encountered resistance from their suppliers
 Lonza pursuing reductions in slip agent-containing materials
throughout the plant
 Some quick-wins by switch part #’s or suppliers, mostly long term
goals

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Sep-14
Impact
 Solubility, filterability, L/E, and toxicology assessments have shown no
impact to patient safety
 Filtration studies performed by Lonza showed that slip agents will not
pass through a sterile filter in buffers utilized at Lonza, with the
exception of detergent-based buffers
 At-scale evidence supports this. Rings/residue have not been found in
equipment/vessels downstream of the post-buffer prep filtration at
Lonza
 Detergent buffers are commonly used for viral inactivation
 Requires detailed calculations on permissible daily exposure (PDE) to justify
no impact to patient safety

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Sep-14
CIP
 How do the bag & slip agent
manufacturers clean their own
equipment?
 Solvents! Ethanol, IPA, solvents with grit
 Volatile
 Flammable
 Manual wipe-down
 Not translatable to CIP of
biopharmaceutical manufacturing
equipment
 Concentrated detergents
 Translatable to CIP
 Key is dissolving hydrophobic residue in
aqueous solutions

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Sep-14
Initial Plant Cleaning Philosophy
 Previously validated CIP for Buffer Prep vessels
 WFI-only at 80-85 C
 ~30-45 minutes contact
 Legacy rings required mechanical restoration of the vessels
surfaces
 “Enhanced Visual Inspection” by maintenance technicians
proceduralized temporarily. Lonza utilized fluorescent droplight
inspection from manway (200 lux)
 Typical full chemical CIP of CIP100/CIP200 was not 100%
successful in one area of the plant
 Exaggerated concentrations of CIP100/CIP200 were also not 100%
successful
 CIP150 was also not successful

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Sep-14
Buffer Ring CIP “version 1.0”
 ProKlenz Booster (PKB) - Successful
 Hydrogen Peroxide + unique surfactant blend

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Sep-14
Buffer Ring CIP “version 1.0”
 First validated use of ProKlenz Booster at Lonza Portsmouth
 Low alarm limit for wash conductivity chosen as 9.5 mS/cm
 This is a 1.0%+1.0% solution, and was the manufacturer’s
recommended concentration to use
 High alarm limit for wash conductivity chosen as 20.0 mS/cm
 This is a 2.5%+2.5% solution
 Manufacturer recommended not exceeding 3%, as 3% can lead to
SS surface oxidation “tinting” with long term use.
 Conservative approach to tank surface maintenance
 0.5% safety factor from the 3%
 Acid wash with each CIP
 Annual acid wash PM CIP’s (more aggressive) on Buffer Prep tanks

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Sep-14
Buffer Ring CIP “version 1.0”
 ProKlenz Booster is not a drop in replacement for skids/automation
design for addition of one chemical per wash
 Not ideal to pre-mix CIP100 with ProKlenz Booster due to stability of
peroxide at high pH (requires frequent turnover of skid day tank)
 ProKlenz Booster must be added after solution heat-up has occurred
 Due to the high foaming potential of ProKlenz Booster at temperatures
below 50°C
 Foaming reduces the effectiveness of the CIP
 Peroxide would break down during heat-up
 This maximizes concentration of peroxide while in solution with
CIP100, as it is designed to work.
 Cleaning is designed to work with both chemicals together
 Both CIP 100 and ProKlenz Booster added via totalized mass, to mass
set point

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Sep-14
Buffer Ring CIP “version 1.0”
 Detectability considerations

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Sep-14
Buffer Ring CIP “version 1.0”
 Continuous Monitoring & Validation
 Both CIP100 and ProKlenz Booster added by mass,
monitored/alarmed by conductivity
 CIP100 is the primary contributor to the conductivity (and pH),
confirmation of the concentration of ProKlenz Booster is inferred
from the success of the CIP100 confirmation (which occurs just
prior) and the continuous monitoring scheme of the CIP.

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Sep-14
Buffer Ring CIP “version 1.0”
 Pros
 Lonza Portsmouth has been successful in the removal of slip
agents from the buffer prep vessels post-CIP since implementation
of this CIP
 Cons
 Some surface oxidation “tinting” observed on equipment surfaces
 Visible rings post-CIP still occur occasionally (infrequent)
 Slip agents not present in these rings, comprised of inorganic Anti-Block
agents and Polymers. Generally all hydrophobic inorganics
 These post-CIP rings are remediated manually
 Wipes wetted with ambient solution of 5% CIP100 + 5% ProKlenz
Booster
 “Enhanced Visual Inspection” by manufacturing personnel in place as a
result

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Sep-14
Buffer Ring CIP “version 2.0”
 Lonza has an ongoing initiative to develop an alternative to the
currently validated peroxide-based CIP with ProKlenz Booster
 Drivers / Goals
1. Obtain 100% cleaning success
2. Utilizing cleaning agents that do not impact the equipment surface
 Phase 1 - Coupon-scale experiment to determine which variables
are the most impactful, comparing rate of removal of residue
 Cleaning Agent
 Temperature
 Concentration
 Utilized Engineering Firm with cleaning lab

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1
 Chemicals chosen:
 CIP100 + ProKlenz Booster
 Baseline required for direct comparison
 CIP100 + CIP Additive
 CIP Additive = non-ionic surfactant blend only
 Cosa CIP 95
 NaOH-based + surfactant blend
 Three Surrogate residues

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1 Surrogate Soil
 Developed surrogate residue based on historical residue identifications
pre and post-CIP
 Pre-CIP rings – The actual residue being cleaned
 Blend of most common slip agents and inorganic anti-blocks / polymers
 Oleamide, Erucamide, Stearamide – Talc / Polyethylene
 Surrogate characteristics:
 White in appearance
 Fails water break test
 Resistant to cleaning:
 Water @ 60C
 1% CIP100 @ 60C for 20 minutes
 Dry wipe / manual
 Dissolves in cleaning solution instead of coming off in chunks
 Can be reproducibly generated and soiled onto coupon
 Adheres in sufficient quantity to facilitate gravimetric analysis

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1 Surrogate Soil
 Oleamide and Erucamide
 Stearamide Surrogate could not be made
that met all criteria
 Surrogate formulation
 Slip agent dissolved in a solvent plus
small percentage of solids (talc/PE),
prepared and spiked at elevated
temperature
 Air dry ambient until solvent is removed
 Heat treat in autoclave
 Bake in oven to dry condensate
 Formation method far different than at-
scale residue
 Complete cleaning not required to make
relative comparisons of rate of removal

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Sep-14
 Emersion batch could not be
used, as coupons would
become re-soiled upon removal
due to hydrophobic residue
floating on the surface
 Falling film device
 Control flow rate, temp
 Representative worst case of
cascade flow on vessel walls
Buffer Ring CIP “version 2.0”
Phase 1 Apparatus

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1 Results

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1 Results

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Sep-14
Buffer Ring CIP “version 2.0”
Phase 1 Results
 Conclusions from Phase 1 coupon-scale cleaning of surrogate
residue
 Temperature is impactful
 80 C performs better than 60 C
 Chemical concentration is impactful, but not as much as
temperature
 3% performs better than 1%
 CIP 100 + CIP Additive outperforms both CIP 100 + ProKlenz
Booster
 High likelihood that CIP 100 + CIP Additive will perform as well
or better than CIP 100 + ProKlenz Booster

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Sep-14
Next Steps
 Supply Chain:
 Continue to put pressure on suppliers to remove slip agents from
materials/processes.
 Continue CIP development
 Phase 2 - Complete final coupon-scale experiments with residue
generated in same manner as at-scale residue
 Optional Phase - Evaluate additional parameters
 Phase 3 - At-scale trials in ad hoc buffer preparations

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Sep-14
Thank You!