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B&D PDGS 04111
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SEPABEADS® FUNCTIONALISED POLYMERS
FOR CHROMATOGRAPHY
Via Roma, 55 – 20082 Binasco (MI) Italy Phone: 0039.02.900130223 – 221 Fax: 0039.02.90090774 e-mail: marketing@resindion.com - technical@resindion.com
B&D PDGS 04111
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Chap. 1. Introduction 3
Chap. 2. Morphological polymer structure 3
Chap. 3. General characteristics summary 5
Chap. 4. SEPABEADS FP/EB area of application 6
Chap. 5. SEPABEADS FP and EB product line 7
Chap. 6. Chromatographic techniques 13
6.1. Ion exchange chromatography 14
6.2. Affinity chromatography 15
6.3. Hydrophobic interaction chromatography 16
6.4. Size exclusion chromatography 17
Chap. 7. Chromatography: from laboratory to industrial scale 18
Chap. 8. Regulatory support file 20
Chap. 9. Custom-made products 20
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1. INTRODUCTION Resindion is now proposing a wide range of specially activated functionalized polymer beads based on hydrophilic highly porous polymer matrix, which offers a very attractive solution for almost every conceivable work-up problem at large scale chromatography application: SEPABEADS FP and EB SERIES: Available in two particle size ranges:
FP grade for fixed-packed bed column application
EB grade for expanded bed column application
2. MORPHOLOGICAL POLYMER STRUCTURE
CO
O R O CO
CO
O
C O
OR ' X
C OO
R O CO
C OO
R ' X
R ' X
O
OR '
X
O
OR
O
O
X = A c t iv e F u n c t io n a l G r o u p
+
S u s p e n s io n P o ly m e r iz a t io n
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POROSITY
PORE DIAMETER (Ǻ)
STANDARD POROSITY GRADE 800 – 1500
HIGH POROSITY GRADE > 5000
Conventional micro porous support
SEPABEADS FP/EB Non conventional
highly porous support
High mechanical resistance
Very rigid: stable internal morphology in water, in organic media and in “dry state”
Conventional macro porous support
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3. GENERAL CHARACTERISTICS SUMMARY
Chemically stable polymer structure
Incompressible spherical beads
Low swelling tendency in high molar solution and in common solvents
High stability in alkalis, acids and solvents
High mechanosmotic stability
High resistance to micro-organism
Available with different porosity
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4. SEPABEADS FP/EB AREA OF APPLICATION The following simplified scheme is given as a guide for the possible application point in an ideal production process. SEPABEADS FP / EB SEPABEADS EC
PRODUCT FERMENTATION
PURIFICATION
PURIFICATION
PURIFICATION
IMMOBILISATION
ENZYME FERMENTATION
BIOCATALYSIS
FINAL PRODUCT
FINAL PRODUCT
FINAL PRODUCT
FINAL PRODUCT
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5. SEPABEADS FP AND EB PRODUCT LINE
The recent progress of biotechnology greatly depends on the development of biomolecules separation technology, and methods for large-scale capture, purification and polishing are of increasing importance.
Among a number of separation methods, Column Liquid Chromatography (LC) is regarded as the most practical process for large-scale industrial applications, because various separation modes can be employed and automatic operations are easily performed.
To achieve a high resolution at industrial-scale, column-packing materials should be carefully designed.
At present time, Resindion R&D developed polymeric sorbent beads usable for industrial applications of Ion Exchange Chromatography (IEC), Affinity Chromatography (AC), Metal Chelate Affinity Chromatography (IMAC), Hydrophobic Interaction Chromatography (HIC) and Size Exclusion Chromatography (SEC).
It has to be noted that the type of functional group identifies SEPABEADS FP/EB Product line, while the particle size derives from the application technique, either fixed bed (FP product line) or expanded bed (EB product line).
Pressure Drop In Water At 25°C
0
2
4
6
8
10
12
14
16
18
20
0 1 2 3 4 5 6 7 8 9 10
LV (m/h)
Pres
sure
dro
p (m
H2O
/m b
ed) SEPABEADS FP SERIES
SEPABEADS EB SERIES
Due to the perfect spherical form of SEPABEADS FP/EB and to the reduced particle size distribution range (uniformity coefficient: max 1.5), the pressure drop produced by a resin bed is very low, as shown in the figure.
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SEPABEADS FP AND EB PRODUCTS LINE
Particle Size (µm) Basic mechanism of separation Product Name
Functional Group
Total Exchange Capacity FP Line
Fixed Bed EB Line
Expanded Bed IEC MCAC (IMAC) AC HIC SEC
CM Carboxymethyl min. 0.1 meq/ml 90 – 250 150 - 300 X X
SP Sulphopropyl min. 0.1 meq/ml 90 – 250 150 – 300 X
DA Diethylamino min. 0.25 meq/ml 90 – 250 150 – 300 X
QA Quaternary ammonium min. 0.2 meq/ml 90 – 250 150 – 300 X
IDA Iminodiacetic min. 50 µmol Zn++/ml 90 – 250 150 – 300 X
HG Diol - 90 – 250 150 – 300 X
BU Butyl - 90 – 250 150 - 300 X
EP Epoxy min. 70 µmol/ml 90 – 250 150 - 300 X
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6. CHROMATOGRAPHIC TECHNIQUES
1. ION EXCHANGE CHROMATOGRAPHY (IEC)
2. AFFINITY CHROMATOGRAPHY (IMAC)
3. HYDROPHOBIC INTERACTION CHROMATOGRAPHY (HIC)
4. SIZE EXCLUSION CHROMATOGRAPHY (SEC)
On the basis of the hydrophilic nature of the acrylic polymers of SEPABEADS,
different kinds of interactions could be used to perform a chromatographic
separation of a compounds mixture applying different chromatographic
techniques.
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6.1. ION EXCHANGE CHROMATOGRAPHY
Different grade of interaction:
Neat charge
Charge density
Charge distribution
Bound sample selectivity removed by changing salt concentration or pH
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6.2. AFFINITY CHROMATOGRAPHY
Separation is achieved by a reversible interaction between a target molecule and a
specific “ligand” blocked to the matrix through a spacer arm.
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6.3.HYDROPHOBIC INTERACTION CHROMATOGRAPHY
Target molecules are charged on the matrix through a high concentration buffer,
condition that allows hydrophobic interaction between the molecule hydrophobic
functional groups present on the matrix surface. Elution takes places applying a
buffer decreasing concentration.
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6.4.SIZE EXCLUSION CHROMATOGRAPHY
Larger MW: shorter retention
time (exclusion limit)
Smaller MW: longer
retention time
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7. CHROMATOGRAPHY:
FROM LABORATORY TO INDUSTRIAL SCALE SEPABEADS FP and EB products are also available in pre-packed columns or bulks1
PRE-PACKED COLUMNS BULKS SEPABEADS
CQK31S (10 µm) CQK31P (10 and 30 µm) FP/EB-CM
CQK30S (10 µm)2 CQK30P (10 and 30 µm) FP/EB-SP
CQA31S (10 µm) CQA31P (10 and 30 µm) FP/EB-DA
CQK35S (10 µm) CQA35P (10 and 30 µm) FP/EB-QA
CQP30 (10 µm) CQP30P (10 and 30 µm) FP/EB-HG
CQH3BS (10 µm) CQH3BS (10 and 30 µm) FP/EB-BU
1 See separately the MCI-GEL catalogue 2 Available also as Protex DEAE and SP at 5 µm
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PARTICLE SIZE RANGE COMPARATIVE TABLE
SEPABEADS FP
SEPHAROSE FF and XL
TOYOPEARL “5…C”
TOYOPEARL “6…M”
EMD FRACTOGEL
SEPABEADS FP
TOYOPEARL “6…C”
EMD FRACTOPREP
STREAMLINE / SEPHAROSE BIG BEADS
SEPABEADS EB
MCI-GEL
PORE SIZE 300 – 600 Å
PORE SIZE 800 - 1200 Å
SEVERAL PORE SIZE
0 10 20 30 40 50 90 100 150 165 250 300 PARTICLE SIZE (µm)
B&D PDGS 04111
Neither this publication, nor any products in this brochure shall be construed as recommending the infringement of any patent, nor extending any license, express or implied, nor assuming any liability under any issued or pending patent. The data presented have been carefully compiled from our records which we believe to be accurate and reliable. We make, however, no warranties or representations with respect hereto, nor is freedom from any patent to be inferred
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8. REGULATORY SUPPORT FILE
RDN commitment to the quality of SEPABEADS products is ruled by ISO9001:2000 as
part of RDN corporate strategy for the highest customer satisfaction degree.
The monomers applied to manufacture SEPABEADS comply with the Resolution
AP(97)1, which rules the use of certain products application in the Food processes.
As Regulatory support file, for each standard product, is released the following
documentation:
• product description;
• material safety data sheet;
• storage conditions and limitations (if any);
• standard specification;
• detailed quality control procedures;
• pre-conditioning, regeneration and standard cleaning in place procedure;
• lot-to-lot certificate of analysis.
9. CUSTOM-MADE PRODUCTS
→ special quality control testing;
→ special Regulatory support studies;
→ beads modification (size and porosity);
→ special ligand development
are offered under request.
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