U.S. patent application number 09/834048 was filed with the patent office on 2002-12-12 for method and apparatus for fractional separation of proteins from plant material.
Invention is credited to Barrilleaux, Adam, Irvine, James E., Mirkov, T. Erik, Monclin, Jean P., Moonan, Francis.
Application Number | 20020187206 09/834048 |
Document ID | / |
Family ID | 22724055 |
Filed Date | 2002-12-12 |
United States Patent
Application |
20020187206 |
Kind Code |
A1 |
Mirkov, T. Erik ; et
al. |
December 12, 2002 |
Method and apparatus for fractional separation of proteins from
plant material
Abstract
A method of extracting and purifying recombinant protein(s) from
transgenic plant matter is disclosed. Fractioning of juice that has
been extracted from the plant matter is obtained by using a
multiple stage filtering process that uses multiple stages of
decreasing porosity (preferably screening) followed by preferably
membrane type filters, ion exchange, membrane adsorber, and
chromatographic processes.
Inventors: |
Mirkov, T. Erik; (Harlingen,
TX) ; Monclin, Jean P.; (Lafayette, LA) ;
Barrilleaux, Adam; (New Iberia, LA) ; Irvine, James
E.; (South Padre Island, TX) ; Moonan, Francis;
(Fairfax, TX) |
Correspondence
Address: |
GARVEY SMITH NEHRBASS & DOODY, LLC
THREE LAKEWAY CENTER
3838 NORTH CAUSEWAY BLVD., SUITE 3290
METAIRIE
LA
70002
|
Family ID: |
22724055 |
Appl. No.: |
09/834048 |
Filed: |
April 11, 2001 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60196085 |
Apr 11, 2000 |
|
|
|
Current U.S.
Class: |
424/725 ;
435/283.1 |
Current CPC
Class: |
C07K 14/47 20130101;
C12N 15/8216 20130101; A61K 38/00 20130101; C07K 14/415 20130101;
C12N 15/8257 20130101; C12N 15/8279 20130101 |
Class at
Publication: |
424/725 ;
435/283.1 |
International
Class: |
A61K 035/78; C12M
001/00 |
Goverment Interests
[0003] This work was supported by DOD Grant No. DAAG55-97-1-0096.
The government may have rights in this invention.
Claims
1. A method of refining transgenic plant matter to extract protein
matter comprising the steps of: a) providing a transgenic plant
matter feedstock to be processed; b) extracting the plant matter
feedstock to extract pression juice; c) cleaning the pression juice
to remove particulate matter; d) adjusting the pH of the juice to a
pH of at least 4.5; e) transmitting the juice from steps "a"
through "d" to a membrane separation system in order to produce two
fractions, one of the fractions containing a protein of
interest:
2. The method of claim 1 further comprising the step of removing
colorant from the juice with ion exchange.
3. The method of claim 1 wherein there is no fermentation involved
for the product or separation of protein.
4. The method of claim 1 wherein the screen system has a porosity
of between about 60-500 microns.
5. The method of claim 1 wherein the juice pH value is adjusted to
a range of between about 5.2 and 8.3.
6. The method of claim 1 wherein in step "c" there are at least one
filter or multiple screens that include multiple screens having
different porosities.
7. The method of claim 1 wherein the membrane separation system of
step "e" wherein there are a plurality of membranes.
8. The method of claim 1 wherein there are a plurality of membrane
separation stations in step "e" and a plurality of fraction tanks,
one fraction tank for each membrane separation station.
9. The method of claim 7 wherein there are at least three membrane
separation stations.
10. The method of claim 7 wherein each membrane separation station
generates a retentate fraction and a permeate fraction.
11. The method of claim 12 wherein there are a plurality of
fraction tanks and each permeate fraction is transmitted to a
tank.
12. The method of claim 1 wherein the transgenic plant matter is
barley.
13. The method of claim 1 wherein the transgenic plant matter is
corn.
14. The method of claim 1 wherein the transgenic plant matter is
potatoes.
15. The method of claim 1 wherein the transgenic plant matter is
alfalfa.
16. A process for extraction of protein from transgenic plant
matter, comprising the steps of: a) extracting juice from a
feedstock of protein contained in the transgenic plant matter; b)
preliminarily screening said juice to remove particulate matter,
using a plurality of screens spanning a range of porosity of
between 50 and 500 microns porosity; c) treating the juice with
multiple stages of ultra-clarifying filtration of decreasing
porosity, including some ultra-clarifying filtration that includes
membrane separation.
17. The process of claim 16 wherein the step of ultra-clarifying
the juice comprises ultra-filtering the juice with a membrane
having a cutoff that will fractionate proteins having molecular
weight between about 5,000 and 500,000.
18. The process of claim 16 wherein the transgenic plant matter is
barley.
19. The process of claim 16 wherein the transgenic plant matter is
corn.
20. The process of claim 16 wherein the transgenic plant matter is
potatoes.
21. The process of claim 16 wherein the transgenic plant matter is
alfalfa.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is a continuation-in-part of co-pending U.S. patent
application Ser. No. ______, filed Apr. 10, 2001, entitled
"Sugarcane Fractioning System", which is incorporated herein by
reference.
[0002] Priority of U.S. Provisional Patent Application Serial No.
60/196,085, filed Apr. 11, 2000, which is incorporated herein by
reference, is hereby claimed.
REFERENCE TO A "MICROFICHE APPENDIX"
[0004] Not applicable
BACKGROUND OF THE INVENTION
[0005] 1. Field of the Invention
[0006] This invention relates to a novel technology for the
separation of proteins from transgenic plant matter (eg. stalk,
leaf and/or grain) and fractional purification of proteins so that
proteins can be separated and further purified from other
compounds. The invention allows for the rapid separation and
fractional purification of large quantities of proteins. The
present invention more particularly relates to a separation and
fractional purification process that can be applied with any type
of transgenic vegetal plant material (including, eg. plant stalk,
leafy material and/or grain) such as but not limited to cane,
barley, corn, potatoes, alfalfa, used to produce essentially any
category of recombinant protein(s) such as but not limited to
monoclonal antibodies (MAB), lectins, collagens, enzymes, or
therapeutic proteins. During each step or any of the steps of this
novel process, unconventional or conventional laboratory analysis
could be performed in order to monitor the streams and the
concentration of the protein(s) of interest.
[0007] 2. General Background of the Invention
[0008] The extraction of the protein(s) of interest from the
feedstock can be performed through different means that will not be
destructive to the different proteins. The extraction can be
performed through a preparation or comminuting step for size
reduction of the feedstock followed by maceration or leaching
steps. Comminuting of the feedstock will be done with apparatus
such as but not limited to crusher, grinder, and cutting machine.
For example, starting with genetically modified sugarcane stalks
containing the protein(s) of interest, and passing the stalks
through a pressure system such as roller/crusher allows extracting
a liquid in this case also called pression juice, which contains
the protein(s) of interest.
[0009] The present invention relates to processing of plant matter
for the recovery (fractional purification) of high value proteins
so that these proteins can be separated and further purified from
other compounds.
[0010] The present invention enables the rapid separation and
fractional purification of large quantities of proteins. This
process is preferably applied to any type of transgenic vegetable
plant material such as, for example; cane, sugarcane, barley, corn,
potatoes, alfalfa, etc., and can be used to produce essentially any
category of recombinant protein(s) such as, but not limited to,
monoclonal antibodies (MAB), lectins, collagens, enzymes, or
therapeutic proteins. During each step or any of the steps of the
process of the present invention, unconventional or conventional
laboratory analysis could be performed in order to monitor the
streams and the concentration of the protein(s) of interest.
BRIEF SUMMARY OF THE INVENTION
[0011] The present invention provides a process for extracting high
value protein(s) directly from transgenic plant material.
[0012] The extraction of transgenic plant feedstock containing the
protein(s) of interest can be performed through different means
that will not be destructive to the different proteins. Genetically
modified and/or non-genetically modified plants containing the
protein(s) of interest and able to produce a juice when passing
through a pressure system can be processed. For example, starting
with genetically modified plant material containing the protein(s)
of interest, and passing the plant matter through a pressure system
such as roller/crusher allows extracting a liquid, in this case
also called "pression juice", which contains the protein(s) of
interest.
[0013] The extracting pressure system can for example use: (a)
different geometry rollers, (b) a plurality of rollers, (c) water
for further extraction of pression juice, (d) series pressure
system where the plant matter after the first pressure system will
feed a second pressure system in series with the first pressure
system, etc. (e) a buffer solution which will avoid partially or
entirely, oxidation or degradation of some compounds contained in
the pression juice.
[0014] In order to improve the juice extraction of the pressure
system, the plant matter can be previously shredded. Both shredder
and pressure system are preferably part of the extraction step. The
extraction step can work continuously or discontinuously.
Extraction could also be performed through a leaching process such
as diffusion.
[0015] Following the extraction step, the pression juice is
transmitted, either by gravity or by means of pumping to a
screening system that is preferably composed of one or several
screening steps. For example, a three step screening system can be
used that is comprised of: (a) a first screening step that removes
selected particulate, eg. matter larger than about 500 microns to
1000 microns, (b) the second screening step can be used for
removing particulate size larger than about 150 microns to 250
microns, (c) the third screening step can be used to remove
particulate size larger than about 10 to 60 microns.
[0016] The screening system can include, for example, screens that
are stationary, vibrating, rotary or any combination of these types
of screens. Screens could also be self-cleaning units. The screened
juice is recovered for further processing and the reject is
discarded or sent to alternate processing. Press filter(s) or other
filtering devices such as pressure filters could be used as an
option to the screening step.
[0017] The screened juice is transferred to a receiving/mixing tank
where its pH is adjusted to a value that is preferably in the range
of between about 5.2 to 8.3, accordingly to the protein(s) of
interest. The tank could be equipped with a low shear rate-mixing
device. The tank is preferably designed to control the temperature
of the juice to a value between about 4.degree. Celsius to
70.degree. Celsius.
[0018] The juice from the receiving/mixing tank is transmitted (eg.
pumped) at constant flow into a first membrane separation system.
This first system performs the separation of suspended solids with
a size larger than between about 0.1 to 0.2 microns. The clean
juice contains the protein(s) of interest. This clean juice or
"first permeate fraction" is sent to a receiving tank before
transmission to the next method step.
[0019] The membrane reject or first retentate fraction is discarded
or sent to alternate processing. The first retentate fraction
contains contaminants such, as but not limited to: dextrans, waxes,
bagacillo, bacterias, yeast, and suspended solids that are
typically larger than about 0.1 to 0.2 microns. Membranes can be of
different types, materials and configurations. As an example,
hollow fiber polymeric membranes can be used. However, composite
membranes can be used as well as inorganic (for example, ceramic
and coated stainless steel tube membranes) and polymeric membranes
with different, selected configurations.
[0020] The first membrane separation system can be comprised of a
single or several membranes working in parallel or in series.
Operating temperature is preferably in the range of between about
4.degree. Celsius to 70.degree. Celsius. Fluxes obtained are
preferably in the range of between about 15 to 160 gfd (gallon per
square foot per day) at different trans-membrane pressures. During
this step some properties of the membrane such as hydrophilicity
can enhance the separation process.
[0021] The permeate (also called clean fraction from the first step
membrane) is collected into a tank called first permeate tank.
[0022] The product from the first fraction tank is used to feed at
preferably constant flow, the second membrane separation system.
This second membrane separation system performs the separation of
particulate larger than between about 0.01 to 0.05 microns. The
permeate fraction is collected into a tank called the second
fraction tank. The retentate fraction is collected into a tank
called second retentate tank. According to its (their) molecular
size(s), the protein(s) of interest could be in either the second
retentate fraction or the second permeate fraction.
[0023] Membranes can be of different types, materials and
configurations. Hollow fiber polymeric membranes can be used.
However, composite membranes can be used as well as inorganic
(ceramic and coated stainless steel tube membranes) and polymeric
membranes all of them with arrangement including hollow fiber,
spiral, plate and tubular module configurations.
[0024] The second membrane separation system can be composed of a
single or several membranes working in parallel or in series.
Operating temperature is preferably in the range of between about
4.degree. Celsius to 70.degree. Celsius. Fluxes can be in the range
of between about 5 to 80 gfd (gallon per square foot per day) at
different transmembrane pressures. The system can be hydraulically
designed in order not to exceed a shear rate of 10,000 sec.sup.-1.
During this step some properties of the membrane such as
hydrophilicity can enhance the separation process. The discarded
fraction is sent to alternate processing.
[0025] The fraction containing the protein(s) of interest, either
the second permeate fraction or the second retentate fraction is
collected into a second fraction tank. From the second fraction
tank, the second fraction is transmitted (eg. pumped) at preferably
constant flow into the third membrane separation system, which has
cut size of about 5,000 to 80,000 molecular weight.
[0026] The membranes used in the third separation system can be
made of different material with different shape and configuration.
Membranes can be of different types, materials and configurations.
The membrane used can be a flat plate configuration, often referred
to as "cassettes". However, hollow fiber and spiral wound membranes
could also be used. Different materials either regenerated
cellulose or polyethersulfone membranes can be used. Other
materials that could be used such as polymeric membranes with
arrangement including hollow fiber, spiral, plate or tubular module
configurations.
[0027] The third membrane separation system can be comprised of a
single or several membranes working in parallel or in series.
Operating temperature is preferably in the range of between about
4.degree. Celsius to 70.degree. Celsius. Fluxes can be in the range
of about 0.1 to 30 gfd (gallon per square foot per day) at
different transmembrane pressures. The system can be hydraulically
designed in order not to exceed a shear rate 10,000 sec.sup.-1.
[0028] The third membrane separation system produces two fractions:
(a) the third permeate fraction and (b) the third retentate
fraction. The protein(s) of interest is (are) in one of these two
fractions. The discarded fraction is sent to alternate
processing.
[0029] The fraction containing the protein(s) of interest is
collected into a third fraction tank prior to any further treatment
step during the purification process. The third fraction tank is a
receiving/mixing tank where the pH of the fraction is adjusted to a
value in the range of between about 5.2 to 8.3, accordingly to the
protein(s) of interest. The third fraction tank can be equipped
with a low shear rate-mixing device. The third fraction tank can
also be temperature controlled to maintain the temperature of the
juice to a value between about 4.degree. Celsius to 70.degree.
Celsius.
[0030] The protein fraction of interest after pH adjustment is
transferred (eg pumped) at a rate of about 0.5 to 3.0 beds volume
per hour through an ion exchange column containing a weak anionic
resin with higher affinity (at this pH of about 4.5 tp 8.3,
preferably 5.2 to 8.3) for colorants than any other compounds.
Temperature during this step is maintained at a value between about
4.degree. Celsius to 70.degree. Celsius. Decoloration of the
incoming feed is between about 25% and 95%.
[0031] The decolorized fraction containing the protein(s) of
interest is collected into a ion product tank where the pH of the
fraction is adjusted to a value in the range of about 5.2 to 8.3,
accordingly to the protein(s) of interest. The ion product tank
could be equipped with a low shear rate-mixing device. The ion
product tank could also be designed to control the temperature of
the juice to a value between about 4.degree. Celsius to 70.degree.
Celsius. The juice from this ion product tank is transferred (eg.
pumped) at a rate of about 0.1 to 3.0 beds volume per hour through
an ion exchange chromatographic process for further purification.
The ion exchange chromatographic process step produces several
fractions, one of them with higher concentration of the protein(s)
of interest. A membrane adsorber could replace the ion exchange
chromatographic step.
[0032] The resulting fraction containing the protein(s) of interest
is collected into an ion exchange chromatographic receiving/mixing
tank where the pH of the fraction is adjusted to a value in the
range of about 5.2 to 8.3, accordingly to the protein(s) of
interest. The ion exchange chromatographic tank could be equipped
with a low shear rate-mixing device. The ion exchange
chromatographic tank could also be temperature controlled to
maintain the temperature of the juice to a value between about
4.degree. Celsius to 70.degree. Celsius. The fraction of the
protein(s) of interest could be sent to a concentration step such
as a low temperature evaporating system for further concentration
such as flash/freeze dry. The product from the concentration step
(eg. evaporation station) contains the fractionated desired
protein(s) partially purified and concentrated.
BRIEF DESCRIPTION OF THE DRAWINGS
[0033] For a further understanding of the nature, objects, and
advantages of the present invention, reference should be had to the
following detailed description, read in conjunction with the
following drawings, wherein like reference numerals denote like
elements and wherein:
[0034] FIG. 1 is a schematic flow diagram illustrating the
preferred embodiment of the method and apparatus of the present
invention.
DETAILED DESCRIPTION OF THE INVENTION
[0035] FIG. 1 is a schematic diagram of the preferred embodiment of
the apparatus of the present invention, designated generally by the
numeral 10. FIG. 1 also shows the various method or process steps
of the preferred embodiment of the apparatus of the present
invention. Transgenic plant processing system 10 receives plant
feedstock 11 that are treated preliminarily by extraction step 13.
The feedstock 11 can be any type of transgenic plant material such
as eg. cane, sugarcane, barley, corn, potatoes, alfalfa. As used
herein, the terms "transgenic material should be construed to
include any plant material such as leafy material, stalks, grain,
or the like.
[0036] The extraction step 13 can start with genetically modified
plant matter 11 containing the protein(s) of interest, and passing
the stalks through a pressure system such as roller/crusher allows
extracting a liquid in this case also called pression juice 14,
which contains the protein(s) of interest.
[0037] The extracting pressure system or extraction step 13 can
use: (a) different geometry rollers, (b) any quantity of rollers,
(c) water 12 for further extraction of pression juice 14, (d)
series pressure system where the plant matter 11 after the first
pressure system will feed a second pressure system in series with
the first pressure system, etc. (e) a water and/or buffer solution
12 which will avoid partially or entirely oxidation or degradation
of some compounds contained in the pression juice 14.
[0038] In order to improve the juice extraction of the pressure
system, the plant matter 11 can be previously shredded. Both
shredder and pressure system are part of the extraction step 13.
The extraction step 13 can work continuously or
discontinuously.
[0039] Extraction 13 could be also performed through a leaching
process such as diffusion.
[0040] Following the extraction step 13, the pression juice 16
feeds, either by gravity or by means of pumping to a screening
system composed of one or several screening steps 17, 19, 21. For
example, a three steps screening system will be composed of: (a)
the first screening step 17 could remove particulate matter larger
than about 500 microns to 1000 microns, (b) the second screening
step 19 used for particulate size larger than about 150 microns to
250 microns, (c) the third screening step 21 removing particulate
size larger than about 10 to 60 microns.
[0041] Screens 17, 19, 21 could be stationary, vibrating, rotary or
any combination of these types of screens. Screens 17, 19, 21 could
also be self-cleaning units. The screened juice is recovered at
mixing tank 24 for further processing and the reject 18, 20, 22 is
discarded or sent to alternate processing.
[0042] Press filter(s) or other filtering devices such as pressure
filters could be used as an option to the screening step comprised
of screens 17, 19, 21.
[0043] The screened juice is transmitted to receiving/mixing tank
24 where its pH is adjusted to a value preferably in the range of
about 5.2 to 8.3, accordingly to the protein(s) of interest. The
tank 24 can be equipped with a low shear rate-mixing device. The
tank 24 can also be temperature controlled to maintain a
temperature of the juice to a value about 4.degree. Celsius to
70.degree. Celsius.
[0044] The juice from the receiving/mixing tank 24 is transmitted
(eg. pumped) at constant flow into a first membrane separation
system 25. This first membrane separation system 25 performs the
separation of suspended solids with a size larger than about 0.1 to
0.2 microns. The clean juice contains the protein(s) of interest.
This clean juice or first permeate fraction is sent to first
fraction tank 27 before going into the next step. The membrane
reject or first retentate fraction 26 is discarded or sent to
alternate processing.
[0045] The first retentate fraction 26 contains contaminants such
as but not limited to: dextrans, waxes, bagacillo, bacterias,
yeast, and suspended solids larger than 0.2 microns. Membranes that
are used in system 25 can be of different types, materials and
configurations. Hollow fiber polymeric membranes can be used;
however, composite membranes can be used as well as inorganic
(ceramic and coated stainless steel tube membranes) and polymeric
membranes all of them with different configurations. The first
membrane separation system 25 can be comprised of a single or
several membranes working in parallel or in series. Operating
temperature is preferably in the range of about 4.degree. Celsius
to 70.degree. Celsius. Fluxes can be in the range of about 15 to
160 gfd (gallon per square foot per day) at different
trans-membrane pressure. During this step at first membrane
separation system 25 some properties of the membrane such as
hydrophilicity can enhance the separation process. As previously
indicated, the permeate also called clean fraction from the first
step membrane is collected into a tank 27 called first fraction
tank.
[0046] The product from the first fraction tank 27 is used to feed
(at preferably constant flow) the second membrane separation system
28. This system 28 performs the separation of particulate larger
than about 0.01 to 0.05 microns. The permeate fraction 29 is
collected into a tank called second permeate fraction.
[0047] The retentate fraction 30 is collected into a tank called
second retentate tank. Accordingly to its (their) molecular
size(s), the protein(s) of interest can be either into the second
retentate fraction 30 or the second permeate fraction 29. Membranes
in the second membrane system 28 can be of different types,
materials and configurations. Hollow fiber polymeric membranes can
be used: However, composite membranes can be used as well as
inorganic (ceramic and coated stainless steel tube membranes) and
polymeric membranes all of them with arrangement including hollow
fiber, spiral, plate and tubular module configurations. The second
membrane separation system 28 can be composed of a single or
several membranes working in parallel or in series.
[0048] Operating temperature is preferably in the range of a value
about 4.degree. Celsius to 70.degree. Celsius. Fluxes can be in the
range of about 5 to 80 gfd (gallon per square foot per day) at
different transmembrane pressure. The second membrane separation
system 28 is hydraulically designed in order not to exceed a shear
rate of 10,000 sec.sup.-1. During this step some properties of the
membranes 28 such as hydrophilicity can enhance the separation
process. Any discarded fraction can be sent to alternate
processing.
[0049] The fraction containing the protein(s) of interest can be
either the second permeate fraction 29 or the second retentate
fraction 30 and is collected into the second fraction tank 31. From
the second fraction tank 31, the second fraction is transmitted
(eg. pumped) at preferably constant flow into the third membrane
separation system 32, which has cut size of about 5,000 to 80,000
molecular weight.
[0050] The membrane(s) used in the third separation system 32 can
be made of different material with different shape and
configuration. Such membranes can be of different types, materials
and configurations. The membrane(s) can be flat plate
configuration, often referred as cassettes. However, hollow fiber
and spiral wound membranes can be used. Different materials such as
either regenerated cellulose or polyethersulfone membranes can be
used. Other materials can be used such as eg. polymeric membranes
with arrangement including hollow fiber, spiral, plate or tubular
module configurations.
[0051] The third membrane separation system 32 can be comprised of
a single or several membranes working in parallel or in series.
Operating temperature is in the range of a value about 4.degree.
Celsius to 70.degree. Celsius. Fluxes can be in the range of about
0.1 to 30 gfd (gallon per square foot per day) at different
transmembrane pressure. The third membrane separation system 32 can
be hydraulically designed in order no to exceed a shear rate 10,000
sec.sup.-1. The third membrane separation system 32 produces two
fractions: (a) the third permeate fraction 33 and (b) the third
retentate fraction 34. The protein(s) of interest is (are) in one
of these two fractions 33, 34. Any discarded fraction can be sent
to alternate processing.
[0052] The fraction containing the protein(s) of interest is
collected into the third fraction tank 35 prior to any further
treatment step during the purification process. The third fraction
tank 35 is preferably a receiving/mixing tank where the pH of the
fraction is adjusted to a value in the range of about 5.2 to 8.3,
accordingly to the protein(s) of interest. The tank 35 can be
equipped with a low shear rate-mixing device. The tank 35 can also
be temperature controlled to maintain the temperature of the
processed juice in the tank 35 to a value about 4.degree. Celsius
to 70.degree. Celsius. The protein fraction of interest after pH
adjustment is transferred (eg. pumped) at a rate of about 0.5 to
3.0 beds volume per hour through an ion exchange column 36
containing a weak anionic resin with higher affinity at this pH of
about 5.2 to 8.3 for colorants than any other compounds.
Temperature during this step at column 36 is maintained at a value
about 4.degree. Celsius to 70.degree. Celsius. Decoloration of the
incoming feed is between about 25% and 95%.
[0053] The decolorized fraction containing the protein(s) of
interest is collected into an ion product receiving/mixing tank 38
where the pH of the fraction is adjusted to a value in the range of
about 5.2 to 8.3, accordingly to the protein(s) of interest. The
tank 38 can be equipped with a low shear rate-mixing device. The
tank 38 can also be temperature controlled to maintain a
temperature of the juice to a value about 4.degree. Celsius to
70.degree. Celsius. The juice from tank 38 is transferred (eg.
pumped) at a rate of about 0.1 to 3.0 beds per volume through an
ion exchange chromatographic process step 39 for further
purification. The ion exchange chromatographic process step 39
produces several fractions, one of them with higher concentration
of the protein(s) of interest. Membrane adsorber could replace the
ion exchange chromatographic step 39.
[0054] The resulting fraction containing the protein(s) of interest
is collected into an ion exchange chromatographic receiving/mixing
tank 41 where the pH of the fraction is adjusted to a value in the
range of about 5.2 to 8.3, accordingly to the protein(s) of
interest. The tank 41 can be equipped with a low shear rate-mixing
device. The tank 41 can also be temperature controlled to maintain
a temperature of the juice to a value about 4.degree. Celsius to
70.degree. Celsius. The fraction of the protein(s) of interest
could be sent to a low temperature concentration step 42 (eg.
evaporating system) for further concentration. Such a concentration
step can be, for example, a flash/freeze dry step. The product from
the concentration step 42 (evaporation station) contains the
fractionated protein(s) 44 partially purified and concentrated.
[0055] The following is a list of suitable parts and materials for
the various elements of the preferred embodiment of the present
invention.
1 PARTS LIST PART NO. DESCRIPTION 10 plant matter fractioning
system 11 plant matter feedstock 12 water/buffer 13 extraction step
14 pression juice 15 reject (bagasse) 16 flowline 17 first screen
18 reject 19 second screen 20 reject 21 third screen 22 reject 23
ph buffer 24 mixing tank 25 first membrane 26 first retentate
fraction 27 first fraction tank 28 second membrane 29 second
permeate tank 30 second retentate fraction 31 second fraction tank
32 third membrane 33 third permeate 34 third retentate 35 third
fraction tank 36 ion exchange 37 rejects 38 ion product tank 39 ion
exchange chrom 40 reject 41 ion exchange chrom tank 42
concentration step 43 condensates 44 partially purified protein
[0056] The foregoing embodiments are presented by way of example
only; the scope of the present invention is to be limited only by
the following claims.
* * * * *