EXPANDED BED techniques like filtration and centrifugation.

It benefits down-

Expanded Bed Adsorption • Affinity and Pre-activated Media

ADSORPTION stream processing by decreasing process time and increasing
is unique in enabling the Capture of biomolecules directly yields, and Captures the product more efficiently than batch-
from unclarified cell suspensions and homogenates, bypassing wise procedures or fluidized bed techniques.
BioProcess

Media
BioProcess Media Dynamic Recommended pH Particle Recommended Base *
capacity size flow rate for expansion matrix
Expanded Bed Adsorption working cleaning
STREAMLINE Q XL Very high 3 - 12 2 - 14 200 µm 300 - 500 cm/h 9
STREAMLINE DEAE Average 2-9 2 - 14 200 µm 200 - 400 cm/h 8
STREAMLINE SP XL Very high 3 - 13 3 - 14 200 µm 300 - 500 cm/h 9
STREAMLINE SP Average 3 - 13 3 - 14 200 µm 200 - 400 cm/h 8
Application area
STREAMLINE Chelating Proteins contaning histidine, 4 - 8,5 3 - 14 200 µm 200 - 400 cm/h 8
cystein or tryptophan residues
STREAMLINE rProtein A ††
IgG 3 - 10 2 - 11 135 µm 200 - 400 cm/h 7

Extreme Load Media.

†† Developed under a license from Repligen Corp. [US Pat. 5,151,350]

AFFINITY chromatography relies on spe-

cific binding of the desired product to ligands
attached to a neutral matrix. Binding and elu-
tion conditions are determined by the nature
of the interaction.
BioProcess
BioProcess Media Application area Recommended pH Particle Specified flow rate † Base *
Media

size Matrix
Affinity media working cleaning
Chelating Sepharose Fast Flow Proteins containing histidine, 4 - 8,5 2 - 14 90 µm 200-400 cm/h, 1
cystein or tryptophan residues 100 kPa, XK 50,
bed height 25 cm
rProtein A Sepharose Fast Flow ††
IgG 3 - 10 2 - 11 90 µm 150-250 cm/h, 1
100 kPa, XK 50,
bed height 25 cm
Protein A Sepharose 4 Fast Flow IgG 3-9 2 - 10 90 µm 150-250 cm/h, 1
100 kPa, XK 50,
bed height 25 cm
Protein G Sepharose 4 Fast Flow IgG, Fab 3-9 2-9 90 µm 150-250 cm/h, 1
100 kPa, XK 50,
bed height 25 cm
Blue Sepharose 6 Fast Flow Interferons, Albumin 4 - 12 3 - 13 90 µm min 400 cm/h, 100kPa, 1
XK 50, bed height 25 cm
Heparin Sepharose 6 Fast Flow Antithrombin III 4 - 12 4 - 13 90 µm 200-400 cm/h, 1
100 kPa, XK 50,
bed height 25 cm
††
Developed under a license from Repligen Corp. [US Pat. 5,151,350]

BioProcess
BioProcess Media Coupling site Recommended pHÆ Particle Specified flow rate † Base *
Media

size matrix
Pre-activated media coupling working cleaning
CNBr-activated Sepharose Primary amino groups 7-9 3 - 11 3 - 11 90 µm 150-250 cm/h, 1
4 Fast Flow 100 kPa, XK 50,
bed height 25 cm
NHS-activated Sepharose Primary amino groups 6-8 3 - 13 3 - 13 90 µm 150-250 cm/h, 1
4 Fast Flow 100 kPa, XK 50,
bed height 25 cm

Æ
Refers to stability of coupling between ligand and
base matrix. Ligands can be less stable.
†
Typical values in our tests
* See Base Matrix Key
 ION EXCHANGE is suitable for rapid sep- emphasize resolution for later purification steps whilst

Ion Exchange Media

arations with large starting volumes and for
concentrating products. Big Beads, XL and Fast
Flow media give high throughput for early
process stages. High Performance media
BioProcess
Media
still showing high capacity. Breakthrough capacities over 150
g BSA/L for XL media.Typical flow rates from 150
to 500 cm/h. SOURCE media offer fast, high resolution sepa-
rations typically for the Polishing stage.

BioProcess Media Dynamic Recommended pH Particle Specified flow rate †

Base *
capacity size matrix
Ion Exchange media working cleaning
Q Sepharose Big Beads Average 2 - 12 2 - 14 200 µm 1200 - 1800 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
SP Sepharose Big Beads Average 4 - 13 3 - 14 200 µm 1200 - 1800 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
Q Sepharose XL Very high 2 - 12 1 - 14 90 µm 400 - 700 cm/h, 100 kPa, 4
XK50, bed height 15 cm
SP Sepharose XL Very high 4 - 13 3 - 14 90 µm 400 - 700 cm/h, 100 kPa, 4
XK50, bed height 15 cm
Q Sepharose Fast Flow Average 2 - 12 1 - 14 90 µm 400 - 700 cm/h, 100 kPa, 1
XK50, bed height 15 cm
DEAE Sepharose Fast Flow Average 2-9 1 - 14 90 µm 300 - 600 cm/h, 100 kPa, 1
XK50, bed height 15 cm
ANX Sepharose 4 Fast Flow (low sub)** Average 2 - 12 1 - 14 90 µm min 150 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
ANX Sepharose 4 Fast Flow (high sub)** Average 2 - 12 1 - 14 90 µm min 200 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
SP Sepharose Fast Flow Average 4 - 13 3 - 14 90 µm 400 - 700 cm/h, 100 kPa, 1
XK50, bed height 15 cm
CM Sepharose Fast Flow Average 4 - 13 2 - 14 90 µm 300 - 600 cm/h, 100 kPa, 1
XK50, bed height 15 cm
Q Sepharose High Performance High 2 - 12 1 - 14 34 µm min 75 cm/h, 250 kPa, 1
BioPilot 60, bed height 30 cm
SOURCE 30Q High 2 - 12 1 - 14 Monosized 2 000 cm/h, 1 MPa, 5
30 µm FineLine 100, bed height 10 cm
SOURCE 15Q High 2 - 12 1 - 14 Monosized 400 cm/h, 1 MPa, 5
15 µm FineLine 100, bed height 10 cm
SP Sepharose High Performance High 4 - 13 3 - 14 34 µm min 100 cm/h, 250 kPa, 1
BioPilot 60, bed height 30 cm
CM Sepharose High Performance High 4 - 13 3 - 14 34 µm min 100 cm/h, 250 kPa, 1
BioPilot 60, bed height 30 cm
SOURCE 30S High 2 - 12 1 - 14 Monosized 2000 cm/h, 1 MPa, 5
30 µm FineLine 100, bed height 10 cm
SOURCE 15S High 2 - 12 1 - 14 Monosized 400 cm/h, 1 MPa, 5
15 µm FineLine 100, bed height 10 cm
Extreme Load Media. †
Typical values in our tests
** Unique Selectivity, Anion exchanger.
* See Base Matrix Key
 REVERSED PHASE (RPC) exploits hydrophobic

Reversed Phase Media & Hydrophobic Interaction

interactions in aqueous/organic solvent mixtures and
is typically used to separate closely similar molecules
during Polishing.
BioProcess
Media
BioProcess Media Resolving power Recommended pH Particle Specified flow rate † Base *
size matrix
Reversed phase media working cleaning
SOURCE 15RPC Very high 1 - 12 1 - 14 Monosized 400 cm/h, 1 Mpa, 5
15 µm FineLINE 100, bed height 10 cm

†
Typical values in our tests
* See Base Matrix Key

HYDROPHOBIC INTERACTION (HIC)

separates proteins and peptides by the strength
of their interaction with hydrophobic ligands strength is lowered. Typical dynamic capacity is 10-15 g
attached to an uncharged matrix. Samples bind BSA/L medium, and typical flow rates range from 50 to 300
at high ionic strength and are eluted as the ionic cm/h.

BioProcess
Media

BioProcess Media Hydrophobicity Recommended pH Particle Specified flow rate † Base *

size matrix
Hydrophobic media working cleaning
Octyl Sepharose 4 Fast Flow Very high 3 - 13 2 - 14 90 µm 150 - 250 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
Butyl Sepharose 4 Fast Flow High 3 - 13 2 - 14 90 µm min 150 cm/h, 100 kPa, 1
XK 50, bed height 25 cm
Phenyl Sepharose 6 Fast Flow Moderate 3 - 13 2 - 14 90 µm 200 - 400 cm/h, 100 kPa, 1
(high sub) XK 50, bed height 25 cm
Phenyl Sepharose 6 Fast Flow Moderate 3 - 13 2 - 14 90 µm 200 - 400 cm/h, 100 kPa, 1
(low sub) XK 50, bed height 25 cm
Phenyl Sepharose Moderate 3 - 13 2 - 14 34 µm min 150 cm/h, 300 kPa, 1
High Performance BioPilot 60, bed height 30 cm
SOURCE 15PHE High 2 - 12 1 - 14 Monosized 400 cm/h, 1 Mpa, 5
15 µm FineLINE 100, bed height 10 cm
SOURCE 15ISO Moderate 2 - 12 1 - 14 Monosized 400 cm/h, 1 Mpa, 5
15 µm FineLINE 100, bed height 10 cm
SOURCE 15ETH Low 2 - 12 1 - 14 Monosized 400 cm/h, 1 Mpa, 5
15 µm FineLINE 100, bed height 10 cm
†
Hydrophobic Interaction media display different selectivities Typical values in our tests
depending on the hydrophobic group. Even ligand density * See Base Matrix Key

plays a role for selectivity. The best way to get started

in the lab is to use the HIC selection kit. A HIC kit
is also available for testing SOURCE media.
 GEL FILTRATION separates macromolecules Flow rates and column dimensions can be varied over

Gel Filtration Media & Desalting Media

on the basis of size. Sample loads are typically up a considerable range: lower flow rates and greater bed
to 8% of bed volume for high resolution, but heights give better resolution.
may be increased for higher productivity where
resolution permits.
BioProcess
BioProcess Media Selectivity Load Theoretical Recommended Particle Recommended Base *
Media

Globular proteins plates pH size flow rate matrix

(MW)
DNA Ex limit
Gel filtration media % of Vc m-1 working cleaning
Superdex 30 prep grade Up to 10 000 4-8 > 10 000 3 - 12 1 - 14 34 µm 10 - 30 cm/h 3
Superdex 75 prep grade 3 000 - 70 000 4-8 > 10 000 3 - 12 1 - 14 34 µm 10 - 30 cm/h 3
Superdex 200 prep grade 10 000 - 600 000 4-8 > 10 000 3 - 12 1 - 14 34 µm 10 - 30 cm/h 3
Sephacryl S-100 1 000 - 100 000 4-6 4 000 3 - 11 2 - 13 50 µm 10 - 30 cm/h 2
High Resolution
Sephacryl S-200 5 000 - 250 000 4-6 4 000 3 - 11 2 - 13 50 µm 10 - 30 cm/h 2
High Resolution 30 base pairs
Sephacryl S-300 10 000 - 1 500 000 4-6 4 000 3 - 11 2 - 13 50 µm 10 - 30 cm/h 2
High Resolution 118 base pairs
Sephacryl S-400 20 000 - 8 000 000 4-6 4 000 3 - 11 2 - 13 50 µm 10 - 30 cm/h 2
High Resolution 271 base pairs
Sephacryl S-500 1 078 base pairs 4-6 4 000 3 - 11 2 - 13 50 µm 10 - 30 cm/h 2
High Resolution
Sepharose 4 Fast Flow Up to 30 000 000 2-4 3 000 3 - 12 2 - 14 90 µm 5 - 10 cm/h 1
Sepharose 6 Fast Flow Up to 4 000 000 2-4 3 000 3 - 12 2 - 14 90 µm 5 - 10 cm/h 1

DESALTING is an extremely useful technique for prepar-

ing a sample or linking steps. Sample volumes
can be 30% of bed volume.
BioProcess
BioProcess media Sample Exclusion Load Recommended Recommended Base *
Media

characteristics limit pH flow rate matrix

Desalting media viscosity (MW) % of Vc working cleaning
Sephadex G-25 Coarse High 5 000 10 - 15 2 - 13 2 - 13 50 - 250 cm/h 6
Sephadex G-25 Medium Medium 5 000 15 - 20 2 - 13 2 - 13 50 - 250 cm/h 6
Sephadex G-25 Fine Low 5 000 20 - 25 2 - 13 2 - 13 50 - 250 cm/h 6
Sephadex G-25 Superfine Low 5 000 25 - 30 2 - 13 2 - 13 50 - 250 cm/h 6

* See Base Matrix Key

COLUMN FLOW RATES

Column Flow Rates and Base Matrix Key

I know my linear velocity (LFR, cm/h) and I want to know what volumetric flow rate (VFR, L/h) I can get with different column diameters (D, mm)
VFR, L/h
LFR, cm/h
D,mm 5 10 25 50 75 100 150 200 300 400 500 750 1 000 2 000 Column
16 0,01 0,02 0,05 0,10 0,15 0,20 0,30 0,40 0,60 0,80 1,0 1,5 2,0 4,0 XK
25 0,02 0,05 0,12 0,25 0,37 0,49 0,74 0,98 1,5 2,0 2,5 3,7 4,9 9,8 STREAMLINE
26 0,03 0,05 0,13 0,27 0,40 0,53 0,80 1,06 1,6 2,1 2,7 4,0 5,3 10,6 XK
50 0,10 0,20 0,49 0,98 1,5 2,0 2,9 3,9 5,9 7,9 10 15 20 39 STREAMLINE, XK
70 0,19 0,38 0,96 1,92 2,9 3,8 5,8 7,7 11,5 15,4 19 29 38 77 INdEX
100 0,39 0,79 2,0 3,9 5,9 7,9 12 16 24 31 39 59 79 157 BPG, INdEX
140 0,8 1,5 3,8 7,7 12 15 23 31 46 62 77 115 154 308 BPG, INdEX
180 1,3 2,5 6,4 13 19 25 38 51 76 102 127 191 254 509 CHROMAFLOW
200 1,6 3,1 7,9 16 24 31 47 63 94 126 157 236 314 628 BPG, INdEX
280 3,1 6,2 15,4 31 46 62 92 123 185 246 308 462 615 1 231 CHROMAFLOW
300 3,5 7,1 18 35 53 71 106 141 212 283 353 530 707 1 413 BPG
400 6 13 31 63 94 126 188 251 377 502 628 942 1 256 2 512 CHROMAFLOW
450 8 16 40 79 119 159 238 318 477 636 795 1 192 1 590 3 179 BPG
500 10 20 49 98 147 196 294 393 589 785 981 1 472 1 963 3 925 CHROMAFLOW
600 14 28 71 141 212 283 424 565 848 1 130 1 413 2 120 2 826 5 652 CHROMAFLOW
800 25 50 126 251 377 502 754 1 005 1 507 2 010 2 512 3 768 5 024 10 048 CHROMAFLOW
1000 39 79 196 393 589 785 1 178 1 570 2 355 3 140 3 925 5 888 7 850 15 700 CHROMAFLOW
1200 57 113 283 565 848 1 130 1 696 2 261 3 391 4 522 5 652 8 478 11 304 22 608 CHROMAFLOW
1600 100 201 502 1 005 1 507 2 010 3 014 4 019 6 029 8 038 10 048 15 072 20 096 40 192 CHROMAFLOW
2000 157 314 785 1 570 2 355 3 140 4 710 6 280 9 420 12 560 15 700 23 550 31 400 62 800 CHROMAFLOW

I know my volumetric flow rate (VFR, l/h) and I want to know at what linear velocity (LFR, cm/h) I should develop my process for
different column diameters (D, mm).
LFR, cm/h
VFR, L/h
D, mm 1 5 10 50 100 150 200 250 500 1 000 2 500 5 000 10 000 25 000 Column
16 498 XK
25 204 1 019 STREAMLINE
26 188 942 1 884 XK
50 51 255 510 STREAMLINE, XK
70 26 130 260 1 300 INdEX
100 13 64 127 637 1 274 1 911 BPG, INdEX
140 6 32 65 325 650 975 1 300 1 625 BPG, INdEX
180 4 20 39 197 393 590 786 983 1 966 CHROMAFLOW
200 3 16 32 159 318 478 637 796 1 592 BPG, INdEX
280 2 8 16 81 162 244 325 406 812 1 625 CHROMAFLOW
300 1 7 14 71 142 212 283 354 708 1 415 BPG
400 4 8 40 80 119 159 199 398 796 1 990 CHROMAFLOW
450 3 6 31 63 94 126 157 315 629 1 573 BPG
500 3 5 25 51 76 102 127 255 510 1 274 CHROMAFLOW
600 2 4 18 35 53 71 88 177 354 885 1 769 CHROMAFLOW
800 1 2 10 20 30 40 50 100 199 498 995 1 990 CHROMAFLOW
1000 1 6 13 19 25 32 64 127 318 637 1 274 CHROMAFLOW
1200 4 9 13 18 22 44 88 221 442 885 CHROMAFLOW
1600 2 5 7 10 12 25 50 124 249 498 1 244 CHROMAFLOW
2000 2 3 5 6 8 16 32 80 159 318 796 CHROMAFLOW

Key to Base Matrix Construction.

Base Type Base Type
matrix matrix
1 Highly cross-linked agarose 6 Cross-linked dextran
2 Composite dextran/N,N’-methylene bisacrylamide 7 Highly cross-linked agarose with metal alloy core
3 Highly cross-linked agarose with bound cross-linked dextran 8 Highly cross-linked agarose with crystalline quartz core
4 Highly cross-linked agarose with bound dextran 9 Highly cross-linked agarose with crystalline quartz core
5 Polystyrene/divinyl benzene polymer with controlled pore size and bound dextran

What’s my pressure unit in different parts of the world?

100 kPa = 1 bar = 14.7 psi = 0.1 MPa
PRODUCT CODE NUMBERS

Ordering Information
FOR A SELECTION OF PACK SIZES

Ion Exchange media Hydrophobic Interaction media

STREAMLINE Q XL 300 ml 17-5075-01 Octyl Sepharose 4 Fast Flow 1L 17-0946-03
7,5 L 17-5075-02 5L 17-0946-04
STREAMLINE DEAE 300 ml 17-0994-01 Butyl Sepharose 4 Fast Flow 500 ml 17-0980-04
7,5 L 17-0994-02 5L 17-0980-04
STREAMLINE SP XL 300 ml 17-5076-01 Phenyl Sepharose 6 Fast Flow (high sub) 1L 17-0973-03
7,5 L 17-5076-02 5L 17-0973-04
STREAMLINE SP 300 ml 17-0993-01 Phenyl Sepharose 6 Fast Flow (low sub) 1L 17-0965-03
7,5 L 17-0993-02 5L 17-0965-04
Q Sepharose Big Beads 1L 17-0989-03 Phenyl Sepharose High Performance 1L 17-1082-03
10 L 17-0989-05 5L 17-1082-04
SP Sepharose Big Beads 1L 17-0657-03 SOURCE 15PHE 200 ml 17-0147-02
10 L 17-0657-05 1L 17-0147-04
Q Sepharose XL 300 ml 17-5072-01 SOURCE 15ISO 200 ml 17-0148-02
5L 17-5072-04 1L 17-0148-04
SP Sepharose XL 300 ml 17-5073-01 SOURCE 15ETH 200 ml 17-0146-02
5L 17-5073-04 1L 17-0146-04
Q Sepharose Fast Flow 300 ml 17-0510-01
5L 17-0510-04 Gel filtration media
DEAE Sepharose Fast Flow 500 ml 17-0709-01 Superdex 30 prep grade 1L 17-0905-03
10 L 17-0709-05 5L 17-0905-04
ANX Sepharose 4 Fast Flow (low sub) 500 ml 17-1286-01 Superdex 75 prep grade 1L 17-1044-02
Custom Designed Media produced on receipt of order. 5L 17-1286-04 5L 17-1044-04
ANX Sepharose 4 Fast Flow (high sub) 500 ml 17-1287-01 Superdex 200 prep grade 1L 17-1043-02
Custom Designed Media produced on receipt of order. 5L 17-1287-04 5L 17-1043-04
SP Sepharose Fast Flow 300 ml 17-0729-01 Sephacryl S-100 High Resolution 750 ml 17-0612-01
5L 17-0729-04 10 L 17-0612-05
CM Sepharose Fast Flow 500 ml 17-0719-01 Sephacryl S-200 High Resolution 750 ml 17-0584-01
10 L 17-0719-05 10 L 17-0584-05
Q Sepharose High Performance 1L 17-1014-03 Sephacryl S-300 High Resolution 750 ml 17-0599-01
5L 17-1014-04 10 L 17-0599-05
SOURCE 30Q 1L 17-1275-03 Sephacryl S-400 High Resolution 750 ml 17-0609-01
5L 17-1275-04 10 L 17-0609-05
SOURCE 15Q 200 ml 17-0947-05 Sephacryl S-500 High Resolution 750 ml 17-0613-01
1L 17-0947-03 10 L 17-0613-05
SP Sepharose High Performance 1L 17-1087-03 Sepharose 4 Fast Flow 1L 17-0149-01
5L 17-1087-04 10 L 17-0149-05
CM Sepharose High Performance 1L 17-1277-03 Sepharose 6 Fast Flow 1L 17-0159-01
Custom Designed Media produced on receipt of order. 5L 17-1277-04 10 L 17-0159-05
SOURCE 30S 1L 17-1273-03
5L 17-1273-04 Desalting media
SOURCE 15S 200 ml 17-0944-05 Sephadex G-25 Coarse 5 kg 17-0034-03
1L 17-0944-03
Sephadex G-25 Medium 5 kg 17-0033-03
Affinity media
STREAMLINE Chelating 300 ml 17-1280-01 Sephadex G-25 Fine 5 kg 17-0032-01
Custom Designed Media produced on receipt of order. 7,5 L 17-1280-02
STREAMLINE rProtein A 300 ml 17-1281-02 Sephadex G-25 Superfine 5 kg 17-0031-03
5L 17-1281-03
Chelating Sepharose Fast Flow 500 ml 17-0575-02
1L 17-0575-04 Reversed Phase Media
rProtein A Sepharose Fast Flow 1L 17-1279-04 SOURCE 15 RPC 200 ml 17-0727-02
5L 17-1279-05 1L 17-0727-04
Protein A Sepharose 4 Fast Flow 1L 17-0974-03
5L 17-0974-05 Pre-activated media
Protein G Sepharose 4 Fast Flow 500 ml 17-0618-03 CNBr-activated Sepharose 4 Fast Flow 250 g 17-0981-03
5L 17-0618-04 2 kg 17-0981-05
Blue Sepharose 6 Fast Flow 1L 17-0948-03 NHS-activated Sepharose 4 Fast Flow 500 ml 17-0906-02
5L 17-0948-04 5L 17-0906-04
Heparin Sepharose 6 Fast Flow 1L 17-0998-03
5L 17-0998-04

Extreme Load Media.