Biotechnology
Biotechnology
Biotechnology
Introduction
1 .l FERMENTATION BROTH
Sedimentation
Solvent extraction
Filtration from whole cdls Ion exchanpe
solvsnf/d~lrrgent Adsorption
rend&-q membranes, Gel filfrstion
rtc, “leaky’! Affinity method9
Distillation
Membranes
Electrophoresir
Differential freezing
Evaporation
Membrsner
Pmcipitalion
Adsorption. ion exchange, clf%ily. erc
Frocra.ThDwing-cryslalliralion
Mechanical Chemical
Filtration Adsorption
Centrifugation Ion exchange
Evaporation Column chromatography
Crystallization Affinity chromatography
Drying Solvent extraction
Reverse osmosis Electrophoresis
Ultrafiltration Electrodialysis
Example 1 .l
t nrS 1 arc V 1
-- + (1.2)
v = P ( A) 2PW ( x2, 1
Introduction 9
The slope and intercept for the pH 3.8 curve can be used to
obtain the values for rs and r,/W from the information
given. Using the data points (300, 0.7) and (600, 1.82), rs =
-55.73 and r,/W = 99.55.
1 _(y t &gyy
(A ) = nrcV
(1.3)
PW
18n . St2
cp = - (1.4)
2
d* (cc - PL) g w rV
Company Location
Allied Filtration Co. Kingsley, PA
Avery Filter Co. Westwood, NJ
Bird Machine Company South Walpole, MA
Carl C. Brimmekamp & Co. Stamford, CT
Denver Equipment Company Colorado Springs, CO
Dorr-Oliver, Inc. Stamford, CT
EIMCO (Div. of Envirotech) Salt Lake City, UT
B C Hoesch Industries Wharton, NJ
Inlay, Inc. Califon, NJ
Komline-Sanderson, Inc. Peapack, NJ
Lenser America, Inc. Lakewood, NJ
Peterson Filters, Inc. Salt Lake City, UT
R & R Filtration Systems Marietta, GA
Serfilco,‘Ltd. Glenview, IL
Sparkler Filters, Inc. Conroe, TX
D.R. Sperry & Co. North Aurora, IL
Example 1.2
FEED
POROUS LINER
FILTER CAKE
PERFORATED FRAME
>
rl
EFFLUENT m EFFLUENT
FILTER CAKE
FEED
b
Figure 1.3. Centrifuges. (a) Perforated bowl or basket type
centrifuge. (b) Tubular bowl centrifuge. (c) Multi chamber
solid bowl centrifuge. (d) Nozzle disk centrifuge. (e) De-
sludger or intermittent discharge centrifuge. (f) Scroll
centrifuges.
Introduction 13
FEED
EFFLUENT CG
INTERMITTENT
FEED
EFFL
Company Location
Alfa Lava& Inc. Fort Lee, NJ
Amtek, Inc. El Cajon, CA
Baker Perkins, Inc. Saginaw, MI
Barrett Centrifugals Worcester, MA
Bird Machine Company South Walpole, MA
Centrico, Inc. Northvale, NJ
Clinton Centrifuge, Inc. Hatboro, PA
Commercial Filters Lebanon, IN
Dorr-Oliver, Inc. Stamford, CT
Heinkel Filtering Systems South Norwalk, CT
Krauss Filtering Systems, Inc. Charlotte, NC
IEC Division, Damon Corp. Needham, MA
Pennwalt Corp., Stokes Division Warminster, PA
Quality Solids Separation Co. Houston, TX
Robatel, Inc. Pittsfield, MA
Sanborn Associates, Inc. Wrentham, MA
Tema Systems, Inc. Cincinnati, OH
Western States Machine Company Hamilton, OH
Q = UAAT. (1.5)
In this equation, Q is the overall rate of heat transfer, U is
the overall heat transfer coefficient for the system, A is the
evaporator surface area and AT is the difference between the
steam temperature and the liquid temperature leaving the
vapor head for natural circulation equipment. For forced
circulation equipment, AT is replaced with the log mean
temperature difference to correct for the fact that the
temperature difference is not constant along the evaporator
surface. The overall heat transfer coefficient, U, is obtained
from pilot plant data or calculated from the individual heat
transfer coefficients, hi
l/U = C l/hi . (1.6)
These theoretical rates are greatly reduced by the
presence of air in other configurations such as multiple
effect evaporators. These venting problems make it difficult
to translate small scale evaporation equipment data to full
scale equipment.
CONCENTFAIE. IP
l
UC)
I:^
FEED
I
VAPOR
-CONDENSATE
FE)ED PROliUCT
a b
STEAM
PRODUCT
STEAM STEAM
PACERS
INT GASKETS
VAPOR AND -
CONCENTRATE DISCHARGE
TO SEPARATOR
BELT DRIVE
MOTOR
VAPOR NOZZLE
FEED NOZZLE
ROTOR
HEATING JACKET
HEATING JACKET
A: HEATING NOZZLE
B: HEATING NOZZLE
HEATING JACKET
LOWER ROTOR
BOTTOM CONE
GUIDE BUSHING
DISCHARGE NOZZLE
Example 1.3
-38 60 a2 116
Company Location
Alfa-Lava1 Fort Lee, NJ
APV Company, Inc. Tonawanda, NY
Dedert Corp. Olympia Fields, IL
Distillation Engineering Co, Livingston, NJ
Goslin-Birmingham Birmingham, AL
HPD, Inc. Naperville, IL
Industrial Filter and Pump Cicero, IL
The Kontro Co., Inc. Orange, MA
Luwa Corporation Charlotte, NC
Paul Mueller Co. Springfield, MO
Niro Atomizer, Inc. Columbia, MD
Pfaudler Co. Rochester, NY
Swenson Process Equipment Harvey, IL
Unitech, Div. of Graver Co. Union, NJ
Henry Vogt Machine Company Louisville, KY
NUCLEATION
TEMPERATURE (T)
NON-CONDENSABLE
GAS OUTLET
COOL I NG
WATER INLET --c(:
BAROMETRIC
CONDENSER
STEAM
JET
SWIRL BREAKER
CIRCULATING PIPE
CONDENSATE
b
Figure 1.8. (continued)
STEAM INJECTOR
‘VAPOR
CRYSTAL SUSPENSION
FEED
- CRYSTALS
OUT
Company Location
APV CREPACO, Inc. Tonawanda, NY
Aqua-Chem, Inc. Milwaukee, WI
Blaw-Knox F & C E Co. Buffalo, NY
Dedert Corp. Olympia Falls, NY
Goslin-Birmingham Birmingham, AL
HPD Incorporated Nape&Be, IL
Niro Atomizer, Inc. Columbia, MD
Pfaudler Corp. Princeton, NJ
Swenson Division (Whiting Corp.) Harvey, IL
Unitech, Division of Graver Union, NJ
u.A
NV =
XV
‘TH- Ts) (1.7)
NV = f Ko
(yw
- Yc) (1.8)
FEED SCRAPING
MEDIUM TEMPERATURE
LOW TEMPERATURE
VACUUM
CONNECTION
DOOR
\ TRAYS
CONNECTIONS
DISCHARGE VALVE
VACUUM
FEED \ PORT CONNECTION
/
J DISCHARGE VALVE
DILUTE REGloN
I
___
TRANSITION
PHASE BOUNDARY
ZONE_ _
I-
DENSE REGION
FEED STREAN -
- DISTRIBUTOR
HOT AIR
a
FEED FROM HIGH @ESSURE PUMP
l.
DRlED
TPRODUCT
b
Figure 1.13. Dryers. (a) Fluidized bed dryer. (b) Spray
dryer. (c) Flash dryer.
34 Separation and Purification Techniques in Biotechnology
RECYCLE
WET
DRY FEED
PRODUCT
0
AIR
0
” k?cq
HEATER
Company Location
Aeromatic Towaco, NJ
Al Jet Equipment Company Plumsteadville, PA
APV Anhydro, Inc. AtUeboro Falls, MA
C.E. Raymond, Combustion Engineering, Inc. Chicago, IL
Dedert Corp. Olympia Falls, IL
Dorr-Oliver, Inc. Stamford, CT
Glatt Air Techniques, Inc. Ramsey, NJ
Komline-Sanderson Engineering Corporation Peapack, NJ
Krauss Maffei Corp. Charlotte, NC
Luwa Corp., Process Div. Charlotte, NC
Mikropul Corp., Micron Products Division Summit, NJ
Niro Atomizer Columbia, MD
Procedyne Corp. New Brunswick, NJ
Renneburg Div., Hey1 and Patterson Pittsburgh, PA
Swenson Process Equipment Inc. Harvey, IL
Wyssmont Corporation Fort Lee, NJ
FEED _ RET~TATE
MEbi8h4~E
------_---
JI
PERNEATE
b
Figure 1.14. (a) Schematic of a simple filtration system. (b)
[dealized membrane filtration system.
dCm
JS = Jv Cf (1 - a) - D; + (1.9)
2
E r --AP
Jv = (1.10)
g n rx
R =
Js 100
‘- JC (1.11)
v f
Example 1.4
Figure 1.15 (34) shows the flux through an
ultrafiltration membrane in a stirred cell as a function of
transmembrane pressure. Why does the stirrer rpm affect the
3’
UM-IO MEMBRANE
M-50 CELL
0065% PROTEIN (1830 RPM)
B-
3,9!tPROTEIN (1830RPH)
6-
I I I I 1
,344 .G89 1.034 1,378 1.722 2 I67
K1
Js = - (Cf - Cp) (1.12)
x
and
Introduction 39
K2
Jv = - (LIP - An> (1.13)
x
POROUS TUBE
FEED
Ilk PERMEATE
STAINLESS
STEEL
TOP PLATE
ACRYLIC
TOP MANIFOL
RETENTATE
OUTLET
MEMBRANE PLATE
SEPARATOR SHEET
FILTRATE
OUTLET
FILTRATE OUTLET
BOT
TENTATE INLET
ROLL UP TO ASSEHRLE a’
FEED SIDE
/-
PERMEATE FLOW
THROUGH PIEHLIRANE)
-) RETENTATE
PERMEATE
OUT
Company Location
Amicon, Div. of W.R. Grace Danvers, MA
Culligan, Div. of Beatrice Northbrook, IL
Dow Chemical Midland, MI
Du Pont Wilmington, DE
Enka America, Inc. Ashville, NC
Gelman Sciences Ann Arbor, MI
HPD Inc. Naperville, IL
Koch Membrane Systems, Inc. Wilmington, MA
Memtek Corp. Billerica, MA
Millipore Bedford, MA
Monsanto St. Louis, MO
Osmonics Minnetonka, MN
Pall East Hills, NY
Romicon Woburn, MA
Separex Corp. Anaheim, CA
UOP Des Plaines, IL
Vaponics, Inc. Plymouth, MA
2
2.3 RT log KD = vs (6s - ‘+I2 - (h2 - 6s) (1.15)
I I
-
where Vs is the molar volume of the distributing solute, 6,
..
is its solubility parameter and 6, and 6, are the solubility
parameters of the two immiscible solvents.
cOvO
= %”
e= cv
0 0 + cvv” 1 + Kg
(1.16)
8 = (1 -eyl (1.17)
n
log( en)
n = 1 + (1.18)
KD”
log(1 -
1 + KDV )
Introduction 45
0
SOLVENT
AWEOUS
BROTH
1
Feed -
Mixev
I *eparator
1
?--
Solvent enriched
virh product
:R IN
Company Location
Alfa-LavaI/DeLavaI Co. Fort Lee, NJ
Baker Perkins, Inc. Saginaw, MI
Escher B.V. The Hague, Netherlands
Kuhni Switzerland
Liquid Dynamics Co. Hempstead, NC
Luwa A.G. Zurich, Switzerland
Westfalia Separators West Germany
Cell
Concentration Partition Yield Purification
Enzyme (%I Coefficient (%) Factor
From Saccharomyces cerevkine
cY-Glucosidase 30 2.5 95 3.2
Glucose-6-phosphate dehydro-
genase 30 4.1 91 1.8
Alcohol dehydrogenase 30 8.2 96 2.5
Hexokinase 30 92 1.6
From Escherichia coli
Fumarase 25 3.2 93 3.4
Aspartase 25 5.7 96 6.6
Penicillin acylase 20 2.5 90 8.2
From Brevibacterium ammoniagenes
Fumarase 20 3.3 83 7.5
From Candida Boidinii
Formate dehydrogenase 33 4.9 90 2.0
From Leuconostoc species
Glucose-6-phosphate dehydro-
genase 35 6.2 94 1.3
(1.19)
LOADING SLOT
PLASTIC PEGS \
I
DRYING RACK
7\ ‘PAPER STRIPS
1 GLASS RODS
RACK SUPPORT
FEED WICK
PARTITION
BASE SECTION
BAFFLE SYSTEM
SOLUTION
SPACER FRAMES,
ANOTHER END FRAME,
PS =
tll - % (1.20)
1
- %
FERMENTER
LIIJUID-LIQUID
EXTRACTION
ANIMAL FEED
RllPPl
_I. __.._..
FMFNT .
BULK ANTIBIOTIC
HARVESTED mow
4
FILTER OFF 1. NIGER HVCELlUH USING ROTARY VACUUM FILTER
4
bD tA(0,9 TO FILTRATE UNTIL pti 5.8
c
CALCIUH CITRATE
4
,bD H2S04 WHILE AT 607
c
FILTER ON ROTARY VACUUN FILTER TO REWOVE CASOQ
4
ACTIVATED CHARCOAL TO DECOLORIZE
4
CATION AND ANION EXCHANGE RESIN COLWNS
4
EVAPORATE TO POINT OF CRYSTALLIZATION AT 36’C
4
CRVSTALS OF CITRIC IWNOHVDRATE SEPARATED IN CONTINUOUS
CENTRIFUGES
c
DRIERS AT 50 - 60'C
ClJLlURE SUPERNATANT
SUPERNATANT
400 LT. Pti 8.8 CONDUCTIVITY 23,000,S
1
DILUTION
ADD 400 LT. DENIN. Hz0 +ll.DOD pS
1
ACIDIFICATION
ADD GLACIAL ACETIC ACID TO PH 5.2
1
ADSORPTION
ADD SP-SEPHADEX C25 ( 750 G)
STIR 1 HR. SETTLING 2 HR.
1
COLLECTRESIN
PACK INTO AN ADJUSTABLE COLUMN, REE(OVE
UNADSDRBED PROTEIN BY PASSING 0.3 H
AMMONIUM ACETATE Pti 6.0
1
ELUTION
NUCLEASE ELUTED WITH 2 M CNH4)2S04
1
DIALYSIS
OVERNIGHT AGAINST DEMIN. H20
1
CONCENTRATION / CENTRIFUGATION
CH3 HOLLOW FIBER UNIT @#ICON), MOL.
WT. CUT OFF lO,OOO; lO,0OO G FOR 30 MIN.
GEL FILTRATION
SEPHADEX G75; O.OlZ ACETIC ACID + 0.1 N
AMMONIUM ACETATE
1
FREEZE DRYING
Example 1.6
J 4 $
EXTRACTlON OF CENTRIFUGAL POLYETHYLENEIMINE POLYETHYLENEIMINE
PELLET WITH CHAOTROPIC PRECIPITATION PRECIPITATION
AGENT (GuHCc)
1 J
DIALYSIS
h OF CENTRIFUGAL
AMMONIUM SULFATE
PRECIPITATION OF
AMMONIUM
PRECIPITATION
SULFATE
OF
SUPERNATANT SUPERNATANT SUPERNATANT
4 & 4
CYANOGENBROMIDE ANION EXCHANGE DIALYSIS OF PELLET
CLEAVAGE OF DIALYSIS CELLULOSE CHROMA-
J
PRECIPITATE TOGRAPHY OF
IMMUNOADSORBENT
DISSOLVED PELLET
COLUMN ~~NocLONAL
EXTRACTION OF RESIDUE
\1 ANTIBODIES)
WITH GuHCL, CATION EXCHANGE
S-SULFONATION (CELLULOSE
CHROMATOGRAPHY) CATION EXCHANGE
4
DIALYSIS CELLULOSE
1
I 1
AMMONIUM SULFATE
CHROMATOGRAPHY
c PRECIPITATION
PRECIPITATE (B CHAIN)
1
J,
GEL FILTRATION
DEAE CELLULOSE
CHROMATOGRAPHY I
SEPHACRYL s-200
GEL FlLTRATlON
I .
L-~PH 5 PRECIPITATION OF
SUPERNATANT (A CHAIN)
JI
REVERSE PHASE HPLC
IBGERILLIC ACID
BULK ENZYMES
BGNOCLONAL
ANTIGODIES
GLYCEROPHOSPHATE
DEHYDROGENASE
8 IO-‘- LUCIFERASE
;
a
s
G
10’s-
FACTOR VI-1
8 a
10-b - UROKINASE 0
Q
THERAPEUTIC ENZVMS
IO” I I I I 1 I I I 1-I
IO” 10-l IO” IO’ IO’ IO’ IO’ IO’ rob IO7 108 IO9
SELLING PRICE (S/KG)
1.4 REFERENCES
32. Belcher, D.W., Smith, D.A., Cook, E.M., Chem Eng, p 112
(Jan 17, 1977)
36. Li, N.N., Long, R.B., Henley, E.J., Ind Ena Chem,
57(3):18 (1965)
Introduction 63
57. Sodeck, G., Modl, J., Kominek, J., Salzbrunn, G., Process
Biochem, 16(6):9 (1981)