By: James Gillis, Keni Mallinen, Brenna Swift CHEE 450 Design Project.
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Transcript of By: James Gillis, Keni Mallinen, Brenna Swift CHEE 450 Design Project.
PRODUCTION OF HUMAN INSULIN:
CRYSTALLIZATIONBy: James Gillis, Keni Mallinen, Brenna Swift
CHEE 450 Design Project
PRESENTATION OUTLINE• Process Overview
• Insulin Crystallization
• Batch Crystallization Equipment
• Design Parameters
• Production Capacity
• Materials of Construction
• Suppliers and Costing
• Sterilization and Recovery
• Comparison to Alternative Unit Operations
INSULIN CRYSTALLIZATION Majority of pharmaceutical products marketed in crystalline forms
• high purity• facilitates subsequent centrifugation and
drying • improves product’s aesthetics
Insulin separation by precipitation using zinc chloride
Rhombohedral, zinc hexamer crystal structure
Batch crystallizers
Similar to stirred tank reactors
BATCH CRYSTALLIZATION EQUIPMENT
OPERATING PARAMETERS: SUPERPRO Conditions, reagents, and their interaction dictate crystallization
rate, yield, size, morphology, etc.
SuperPro parameters compared to conditions and reagents utilized in previous insulin crystallization procedures
No measure of pH in SuperPro; extent of crystallization set manually at 90%
Component Flow rate (kg/batch)
Mass Comp. (%)
Concentration (g/L)
Stream S-168: Stream from Diafiltration Unit OperationAcetic Acid 2.21909 1.2602 12.653585
Insulin 12.78009 7.2579 72.874061
WFI 161.08573 91.4818 918.535910Stream S-126: Charge Stream for Crystallization Tank
Ammonium Acetate
1.13267 36.1567 361.56697
Zinc Chloride 2.00000 63.8433 638.43303
CRYSTALLIZATION CONDITIONS Crystallization of insulin achieved by precipitation of
insoluble zinc-hexamer in aqueous solution, complemented by temperature decrease
Inlet Protein Concentration Amorphous precipitate forms if concentration is too high
Temperature Crystallization begins at ambient temperature (25oC) Cooling to 5oC further decreases solubility
pH Use ammonium acetate buffer to fix pH at approximately
6.1 Acetate anions also improve crystallization
Metal Ion ConcentrationDivalent metal ions (group 15 elements)
mediate intermolecular contactsZinc chloride- soluble in acetic acid, allows
formation of zinc insulin hexamer (pictured)0.08g zinc/g insulin
Other Potential SolventsAddition of 10% (v/v) acetoneIncrease step growth kinetics of insulin
crystals
CRYSTALLIZATION CONDITIONS
PRODUCTION CAPACITY (SUPERPRO)
Insulin (12.78 kg/batch)
WFI (161.09 kg/batch)
Zinc Chloride (2.00 kg/batch)
INPUTS
Acetic Acid (2.22 kg/batch)
Ammonium Acetate (1.13 kg/batch)
OUTPUTS
Acetic Acid (2.22 kg/batch)
Zinc Chloride (2.00 kg/batch)
Insulin (1.28 kg/batch)
Ammonium Acetate (1.13 kg/batch)
WFI (161.09 kg/batch)
Insulin Crystal (11.50 kg/batch)
Total Mass Flow Rate = 176.09 kg/batchTotal Volumetric Flow Rate = 175.37 L/batch
Assumptions:• Working volume is 75% of total tank volume• Safety factor of 15%• REQUIRED TANK SIZE = 270 L = 71 US gal
BATCH-TIME OPTIMIZATION
Complete economic analysis is necessary to find optimal operating throughput (number of batches)
Capital and operating cost optimization for entire process
Equipment considerations to prevent bottlenecks and downtime (batch and continuous units)
0 200 400 600 800 1000 1200 14000
10
20
30
40
50
60
70
80
90
100
Time (minutes)
Cry
stal
liza
tio
n (
%)
Crystallization extent as a function of time for porcine insulin (adapted from Schlichtkrull, 1958)
MATERIALS OF CONSTRUCTION
304 SS 316 SS Carbon Steel
Acetic Acid D-Severe Effect
B-Good C-Fair
Ammonium Acetate
B-Good A-Excellent Unknown
Zinc Chloride B-Good B-Good D-Severe Effect
Steel provides excellent strength, durability, and ease of cleaning/sterilization
Three types of steel investigated for chemical compatibility with chemicals present in the inlet/outlet streams Carbon Steel
Stainless Steel 304
Stainless Steel 316
Type 316 SS chosen as material of construction for crystallization tank
Chemical Compatibility of Various Materials with Inlet Chemicals
COSTING AND SUPPLIERS Unsuccessful attempts to contact vendors
Capital cost estimate using Matches online utility
270 L stainless steel crystallizer = $ 65,378.76 (2009 CAD)
Approximate dimensions assuming H:D ratio of 3 0.5 m diameter x 1.5 m height
Adjusted 2007 USD cost using CEPCI and exchange rate of 0.80 USD/CAD
Total cost of chemicals per batch: approximately $390 CAD/batch ($576.00 with 10% v/v acetone)
Potential suppliers: Solvias Inc. (Fort Lee, NJ), Paul Mueller Co. (Springfield, MI)
SEPARATION AND STERILIZATION Crystal separation is achieved by basket
centrifugation following V-106
Steam-in-place (SIP) sterilization between batches using saturated steam (15 psi, 121°C)
ALTERNATIVE UNIT OPERATIONS Several options available to obtain unique drug types or zinc free insulin:
Gel filtration chromatography Ultrafiltration Diafiltration
These units can successfully purify insulin, though a crystalline product would not be obtained
Other common crystallization procedures include: Evaporative batch crystallization Adiabatic crystallization Continuously seeded crystallization
Wide variety of reagents available and acceptable for insulin crystallization Use of phosphate & citrate buffers Crystallization of monomeric insulin analogs