Protein Purification by Ion-Exchange Chromatography

List of Commercially Available Weak Anion Exchangers^*

Last updated on January 1, 2013. REACH Devices does not manufacture, does not distribute, and does not promote any ion-exchange resins. We develop, manufacture and sell chromatography detectors and detector/gradient pump systems for FPLC, LPLC and MPLC.

2-2.5

2.5-3

3-5

5-7

7-10

10-11

>11

Weak anion exchanger

DEAE

Diethyl aminoethyl

-O-CH₂-CH₂-NH⁺(CH₂-CH₃)₂

6-9^*

Z=+1

0<Z<1

Z=0

DMAE

Dimethyl aminoethyl

-O-CH₂-CH₂-NH⁺(CH₃)₂

~10

Z=+1

0<Z<1

Z=0

^*See main Ion-Exchange Chromatography page for detailed description.

	Diameter, μm: mean (range)	Material	Exclusion limit, (Pore size)	Ionic capacity, μeq/ml	Binding capacity, mg/ml Lysozyme^E (human IgG)^{ELysozyme: pI ~11, MW = 14.5kDa; Human IgG: pI~7.4-8.6, MW = 150kDa; Bovine COHb: MW = 69kDa.}	Max Pressure, psi	pH stability: long term (short term)	Material Trade name
1	400 (300-500)	Agarose, 4% cross-linked, hardened^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	No data	No data	50^{E1No data about the binding conditions.}	9^{F1By analogy with data for Q-Cellthru Bigbeads Plus in A. Staby, I. H. Jensen, Journal of Chromatography A, 908, 2001, 149–161.}	2-14	DEAE Cellthru BigBead Plus^01Sterogene
2	200 (100-300)	Agarose with a quartz core, 6% cross-linked^{B1Supplier does not provide data about volume changes due to ionic strength or pH.}	4,000kDa	130-210	>40^{E1No data about the binding conditions.}	4	2-12 (2-14)	Streamline DEAE^{02GE Life Sciences (former Amersham biosciences)}
3	124 (70-220)	Dextran, cross-linked^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	30kDa	510-680^{D1Supplier provides ionic capacity data as 3-4 meq/dry gram. Number in the table obtained by applying the experimentally established column packing density of 0.17 dry g/ml ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. )}	(30)^{E2Flow rate of 75 cm/h in 0.05 M Tris, pH 8.3.}	2.5	2-13	DEAE Sephadex A-25^{02GE Life Sciences (former Amersham biosciences)}
4	No data	Cellulose, microgranular preswollen^{B1Supplier does not provide data about volume changes due to ionic strength or pH.}	No data	160-195^{D2Supplier provides ionic capacity data in meq/dry gram: 0.88-1.08 for DE-52, 1.8-2.2 for DE-53 and 1.0 for Express-Ion D. Numbers in the table obtained by applying the experimentally established column packing density: 0.18 for both DE-52 and DE-53 and 0.23 for Express-Ion D ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	130^{E3No flow rate reported, 10 mM phosphate buffer, pH 8.5.}	No data^{F2Flow of 221 cm/h at 1.}	2-9.5	DE 52^{03Whatman (part of GE Healthcare)}
5	No data	Cellulose, microgranular preswollen^{B1Supplier does not provide data about volume changes due to ionic strength or pH.}	No data	325-400^{D2Supplier provides ionic capacity data in meq/dry gram: 0.88-1.08 for DE-52, 1.8-2.2 for DE-53 and 1.0 for Express-Ion D. Numbers in the table obtained by applying the experimentally established column packing density: 0.18 for both DE-52 and DE-53 and 0.23 for Express-Ion D ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	150^{E3No flow rate reported, 10 mM phosphate buffer, pH 8.5.}	No data^{F3Flow of 137 cm/h at 1.}	2-12	DE 53^{03Whatman (part of GE Healthcare)}
6	60-130 fibers	Cellulose, microgranular^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	No data	230^{D2Supplier provides ionic capacity data in meq/dry gram: 0.88-1.08 for DE-52, 1.8-2.2 for DE-53 and 1.0 for Express-Ion D. Numbers in the table obtained by applying the experimentally established column packing density: 0.18 for both DE-52 and DE-53 and 0.23 for Express-Ion D ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	60^{E1No data about the binding conditions.}	>10	2-9^{G1Working pH range.}	Express-Ion D^{03Whatman (part of GE Healthcare)}
7	100 (40–160)	Cellulose, beaded ^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	1,000kDa	110–140	(160)^{E4Drained medium, 0.5 M Tris, pH 8.3.}	No data^{F4Max linear flow rate 50 cm/h.}	2-12	DEAE Sephacel^{02GE Life Sciences (former Amersham biosciences)}
8	100 (90-120)	Methacrylic polymer^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	5,000kDa (1,000Å)	50-110	25^{E5Dynamic at 10 % Breakthrough, 1 mg/mL BSA in 50 mM Tris, pH 8.5.}	43^{F5Flow of 150 -250 cm/h is achieved at 15/}	2-13	Toyopearl DEAE-650C^{04TOSOH Biosep LLC (former TosoHaas)}
9	90 (45–165)	Agarose, 6% crosslinked^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	4,000kDa	130–170	(170)^{E6Flow rate of 300 cm/h, 0.05 M Tris, pH 8.3.}	2	3-12 (2-14)	DEAE Sepharose CL-6B^{02GE Life Sciences (former Amersham biosciences)}
10	90 [67^{A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.}] (45–165)	Agarose, 6% crosslinked^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	4,000kDa	110–160	47^{E7Flow rate of 300 cm/h, 10% breakthrough.}	43	2-12 (2-14)	DEAE Sepharose Fast Flow^{02GE Life Sciences (former Amersham biosciences)}
11	90 (45-165 )	Agarose, 4% crosslinked^{B2, B3Supplier reports that volume changes due to ionic strength or pH are negligible. Contain Diethylaminopropyl group instead of Diethylamiethyl group, providing slightly different selectivity then DEAE Sepharose FF. Pore size distribution of ANX Sepharose 4 Fast Flow (high sub) has been optimized for the separation of large proteins.}	30,000kDa	130-170	43^E7	43	3-13 (2-14)	ANX Sepharose FF (high sub)^{02GE Life Sciences (former Amersham biosciences)}
12	90 (45-165 )	Agarose, 4% crosslinked^{B1, B3Supplier does not provide data about volume changes due to ionic strength or pH. Contain Diethylaminopropyl group instead of Diethylamiethyl group, providing slightly different selectivity then DEAE Sepharose FF. Pore size distribution of ANX Sepharose 4 Fast Flow (high sub) has been optimized for the separation of large proteins.}	No data	60-80	No data	No data^F6	3-13 (2-14)	ANX Sepharose FF (low sub)^{02GE Life Sciences (former Amersham biosciences)}
13	90	Agarose, cross-linked, dextran surface extender^{B1Supplier does not provide data about volume changes due to ionic strength or pH.}	No data	290-350	No data^{E8Surprisingly given not for BSA, but for ovalbumin: 90 mg/ml (Flow rate of 600 cm/h, 10% breakthrough, 0.05 M Tris, pH 8.0).}	43	2-12 (2-14)	Capto DEAE^{02GE Life Sciences (former Amersham biosciences)}
14	(40 –130)	Cellulose, cross-linked, beaded^{B4Supplier reports 5% shrinkage/swelling on change from pure water to 0.5M NaCl.}	30kDa	144^{D3Supplier provides ionic capacity data in meq/dry gram: 0.9 for Cellufine A-200; 1.3 for Cellufine A-500 and 0.8 for Cellufine A-800. Numbers in the table obtained by applying the experimentally established column packing densities: 0.16 dry g/ml for Cellufine A-200; 0.12 dry g/ml for Cellufine A-500 and 0.10 dry g/ml for Cellufine A-800 ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	>80^{E1No data about the binding conditions.}	29	2-12 (1-14)	Cellufine A-200^{05JNC corporation, Japan}
15	(40 –130)	Cellulose, cross-linked, beaded^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	500kDa	156^{D3Supplier provides ionic capacity data in meq/dry gram: 0.9 for Cellufine A-200; 1.3 for Cellufine A-500 and 0.8 for Cellufine A-800. Numbers in the table obtained by applying the experimentally established column packing densities: 0.16 dry g/ml for Cellufine A-200; 0.12 dry g/ml for Cellufine A-500 and 0.10 dry g/ml for Cellufine A-800 ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	>60^{E1No data about the binding conditions.}	29	2-12 (1-14)	Cellufine A-500^{05JNC corporation, Japan}
16	(40 –130)	Cellulose, cross-linked, beaded^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	1,000kDa	80^{D3Supplier provides ionic capacity data in meq/dry gram: 0.9 for Cellufine A-200; 1.3 for Cellufine A-500 and 0.8 for Cellufine A-800. Numbers in the table obtained by applying the experimentally established column packing densities: 0.16 dry g/ml for Cellufine A-200; 0.12 dry g/ml for Cellufine A-500 and 0.10 dry g/ml for Cellufine A-800 ( P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207. ).}	>45^{E1No data about the binding conditions.}	29	2-12 (1-14)	Cellufine A-800^{05JNC corporation, Japan}
17	(40 –130)	Cellulose, cross-linked, dextran surface extender^{B1Supplier does not provide data about volume changes due to ionic strength or pH.}	>1,000kDa	140-250	197^{E9Dynamic at 10 % Breakthrough,}	43	2-12	Cellufine MAX DEAE^{05JNC corporation, Japan}
18	60^{A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.} (40-90)	Methacrylic polymer, cross-linked, proprietary surface extender^{B5Long, linear polymer chains (“tentacles") carry the functional ligands.}	(800Å)	No data	100^{E10At flow rates of up to 600 cm/h.}	116	1-13	Fractogel DEAE (M)^{06EMD Millipore (Merck Millipore)}
19	(40-90)	Methacrylic polymer, cross-linked, proprietary surface extender^{B5Long, linear polymer chains (“tentacles") carry the functional ligands.}	(800Å)	No data	100^{E10At flow rates of up to 600 cm/h.}	116	1-13	Fractogel DMAE (M)^{06EMD Millipore (Merck Millipore)}
20	59^{A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.} (40-90)	Methacrylic polymer^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	5,000kDa (1,000Å)	80-120	25^{E5Dynamic at 10 % Breakthrough, 1 mg/mL BSA in 50 mM Tris, pH 8.5.}	44^{F5Flow of 150 -250 cm/h is achieved at 15/}	2-13	Toyopearl DEAE-650M^{04TOSOH Biosep LLC (former TosoHaas)}
21	50 [59^{A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.}] (40-90)	Ceramic scaffold filled w/ polymerized hydrogel^{B6Supplier reports the complete absence of volume changes due to ionic strength or pH.}	No data	>200	>85^{E11No flow rate reported, 5 mg/mL BSA in 50 mM Tris, pH 8.6.}	1000	2-12 (1-14)	DEAE Ceramic HyperD F^{07PALL Life Sciences (former BioSepra)}
22	50 [54^{A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.}] (25-100)	Methacrylic polymer^{B7Supplier reports <5% shrinkage/swelling on change from pure 0.1 to 1M NaCl.}	(1,000Å)	100-250	30-40^{E12flow rates 700-100 cm/h, 3 mg/mL BSA in 10 mM Tris, pH 7.6.}	102	1-10	Macro-Prep DEAE^08Bio-Rad
23	35	Methacrylic polymer^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	5,000kDa (1,000Å)	80-120	25^{E5Dynamic at 10 % Breakthrough, 1 mg/mL BSA in 50 mM Tris, pH 8.5.}	44^{F5Flow of 150 -250 cm/h is achieved at 15/}	2-13	Toyopearl DEAE-650S^{04TOSOH Biosep LLC (former TosoHaas)}
24	30 (20-40)	Methacrylic polymer^{B2Supplier reports that volume changes due to ionic strength or pH are negligible.}	5,000kDa (1,000Å)	50-110	20-40^{E1No data about the binding conditions.}	290	2-13	TSKgel DEAE-5PW (30µm)^{04TOSOH Biosep LLC (former TosoHaas)}
25	(20-40)	Methacrylic polymer, cross-linked, proprietary surface extender^{B5Long, linear polymer chains (“tentacles") carry the functional ligands.}	(800Å)	No data	100^{E10At flow rates of up to 600 cm/h.}	116	1-13	Fractogel DEAE (S)^{06EMD Millipore (Merck Millipore)}

^A– Experimental value determined by Coulter Counting, in A. Staby, J. Nielsen, J. Krarup et all, Advances in Resins for Ion-Exchange Chromatography, in Advances in Chromatography, Edited by Nelu Grinberg and Eli Grushka, Volume 47, 2009, 193–245.

^B1-Supplier does not provide data about volume changes due to ionic strength or pH. ^B2-Supplier reports that volume changes due to ionic strength or pH are negligible. ^B3-Contain Diethylaminopropyl group instead of Diethylamiethyl group, providing slightly different selectivity then DEAE Sepharose FF. Pore size distribution of ANX Sepharose 4 Fast Flow (high sub) has been optimized for the separation of large proteins. ^B4-Supplier reports 5% shrinkage/swelling on change from pure water to 0.5M NaCl. ^B5-Long, linear polymer chains (“tentacles") carry the functional ligands. ^B6-Supplier reports the complete absence of volume changes due to ionic strength or pH. ^B7-Supplier reports <5% shrinkage/swelling on change from pure 0.1 to 1M NaCl.

^D1-Supplier provides ionic capacity data as 3-4 meq/dry gram. Number in the table obtained by applying the experimentally established column packing density of 0.17 dry g/ml (P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207.) ^D2-Supplier provides ionic capacity data in meq/dry gram: 0.88-1.08 for DE-52, 1.8-2.2 for DE-53 and 1.0 for Express-Ion D. Numbers in the table obtained by applying the experimentally established column packing density: 0.18 for both DE-52 and DE-53 and 0.23 for Express-Ion D (P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207.). ^D3-Supplier provides ionic capacity data in meq/dry gram: 0.9 for Cellufine A-200; 1.3 for Cellufine A-500 and 0.8 for Cellufine A-800. Numbers in the table obtained by applying the experimentally established column packing densities: 0.16 dry g/ml for Cellufine A-200; 0.12 dry g/ml for Cellufine A-500 and 0.10 dry g/ml for Cellufine A-800 (P. Levison, C. Mumford, M. Streater, A. Brandt-Nielsen, N.D. Pathirana, S.E. Badger, J. Chromatography. A, 160, 1991, 207.).

^E-Lysozyme: pI ~11, MW = 14.5kDa; Human IgG: pI~7.4-8.6, MW = 150kDa; Bovine COHb: MW = 69kDa. ^E1-No data about the binding conditions. ^E2-Flow rate of 75 cm/h in 0.05 M Tris, pH 8.3. ^E3-No flow rate reported, 10 mM phosphate buffer, pH 8.5. ^E4-Drained medium, 0.5 M Tris, pH 8.3. ^E5-Dynamic at 10 % Breakthrough, 1 mg/mL BSA in 50 mM Tris, pH 8.5. ^E6-Flow rate of 300 cm/h, 0.05 M Tris, pH 8.3. ^E7-Flow rate of 300 cm/h, 10% breakthrough. ^E8-Surprisingly given not for BSA, but for ovalbumin: 90 mg/ml (Flow rate of 600 cm/h, 10% breakthrough, 0.05 M Tris, pH 8.0). ^E9-Dynamic at 10 % Breakthrough, ^E10-At flow rates of up to 600 cm/h. ^E11-No flow rate reported, 5 mg/mL BSA in 50 mM Tris, pH 8.6. ^E12-flow rates 700-100 cm/h, 3 mg/mL BSA in 10 mM Tris, pH 7.6.

^F1-By analogy with data for Q-Cellthru Bigbeads Plus in A. Staby, I. H. Jensen, Journal of Chromatography A, 908, 2001, 149–161. ^F2-Flow of 221 cm/h at 1. ^F3-Flow of 137 cm/h at 1. ^F4-Max linear flow rate 50 cm/h. ^F5-Value provider in the Tosoh website. Matarial data sheers (downloaded from the same website) give max pressure value ob 102. ^F5-Flow of 150 -250 cm/h is achieved at 15/

^G1-Working pH range.

⁰¹-Sterogene ⁰²-GE Life Sciences (former Amersham biosciences) ⁰³-Whatman (part of GE Healthcare) ⁰⁴-TOSOH Biosep LLC (former TosoHaas) ⁰⁵-JNC corporation, Japan ⁰⁶-EMD Millipore (Merck Millipore) ⁰⁷-PALL Life Sciences (former BioSepra) ⁰⁸-Bio-Rad

Protein Purification by Ion-Exchange Chromatography

List of Commercially Available Weak Anion Exchangers*

List of Commercially Available Weak Anion Exchangers^*