Macro-Prep 25 Q Anion Exchange Media

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Overview

Property	DEAE	High Q	High S	CM
Type of support	Weak anion	Strong anion	Strong cation	Weak cation
Functional ligand	–N⁺(C₂H₅)₂	–N⁺(CH₃)₃	–SO₃^-	–COO^-
Ionic capacity	175 ±75 µeq/ml	400 ±75 µeq/ml	160 ±40 µeq/ml	210 ±40 µeq/ml
Typcial binding capacity	35 mg BSA/ml	40 mg BSA/ml	70 mg IgG*/ml	35 mg hemoglobin/ml
Counterion	Cl^-	Cl^-	NA⁺	NA⁺
Nominal particle size	50 µm	50 µm	50 µm	50 µm
Nominal pore size	1,000 Å	1,000 Å	1,000 Å	1,000 Å
Recommended maximum linear flow rate	3,000 cm/hr	3,000 cm/hr	3,000 cm/hr	3,000 cm/hr
Chemical stability
1% SDS, 24 hr	Yes	Yes	Yes	Yes
6 M guanidine-HCl, 24 hr	Yes	Yes	Yes	Yes
Volume changes
pH 4–10	<1%	<1%	<3%	<1%
0.1–1.0 M NaCl	<5%	<5%	<9%	<4%
Autoclavability (121°C, 80 min)	Yes	Yes	Yes	Yes
pH stability	1–10	1–10	1–12	1–12
Antimicrobial agent	20% ethanol	20% ethanol	20% ethanol	20% ethanol
Regeneration	1 M NaCl	1 M NaCl	1 M NaCl	1 M NaCl

* Human IgG.

Selection Guide

Product Selection by Technique

		Scale			Packaging Format
Characteristics	Product Type	Analytical	Pilot/Preparative	Process	Prepacked	Bottled
Ion Exchange Chromatography
Separation by charge	UNOsphere™ media					UNOsphere Q, S, Rapid S
General biomolecules purification	Macro-Prep^® media					Macro-Prep High Q, DEAE, High S, CM
Applicable to all antibody purifications	Macro-Prep 25 media					Macro-Prep 25 Q, 25 S
	UNO^® columns				UNO Q, S
	Bio-Scale™ Mini Cartridges				UNOsphere Q, S
High capacity					Macro-Prep High Q, S
Good capture technique	AG^® resins					AG
	Chelex^® resins					Chelex
	Bio-Rex^® resins					Bio-Rex
Hydroxyapatite and Fluoroapatite Chromatography
Unique selectivity	CHT™ 80 µm support					CHT, Types I and II
Alternative to protein A for antibody purification	CHT 40 µm support					CHT, Types I and II
	CHT 20 µm support					CHT, Types I and II
	CFT 40 µm support					CFT, Types I and II
Removal of leached protein A, aggregates, endotoxins, and DNA	Bio-Gel^® HT and HTP media					Bio-Gel HT, HTP
	Bio-Scale columns				CHT Type I (10 µm)
					CHT Type II (20 µm)
Affinity Chromatography
Purification of His- and GST- tagged proteins	Profinity™ IMAC and GST media				Bio-Scale Mini	Profinity IMAC
Separation by biological interaction	Affi-Gel resins					Affi-Gel^® Protein A*
						Affi-Gel Blue*
						DEAE Affi-Gel Blue*
						CM Affi-Gel Blue*
High selectivity						Affi-Gel boronate, heparin**
High purity in a single step						Affi-Gel Hz, 10, 15**
						Affi-Gel 102**
Excellent capture technique	Profinity epoxide media					Profinity epoxide**
	Affi-Prep media					Affi-Prep protein A*
						Affi-Prep polymyxin*
	Bio-Scale Mini cartridges				Profinity IMAC and GST
	Econo-Pac^® columns				Affi-Gel Blue
					DEAE Affi-Gel Blue
					Affi-Gel protein A
Size Exclusion Chromatography
Separation by size	Bio-Gel P media					Bio-Gel P
MW determination	Bio-Beads™ S-X media					Bio-Beads S-X
Applicable to all antibody purifications	Bio-Sil^® and Bio-Silect™ columns				Bio-Sil, Bio-Silect
Desalting or buffer exchange	Bio-Scale Mini cartridges				Bio-Gel P-6
High resolution	Econo-Pac columns				Econo-Pac 10DG
Hydrophobic Interaction Chromatography
Separation by hydrophobicity	Macro-Prep HIC media					Macro-Prep t-butly
						Macro-Prep methyl

* Ready to use.
** Activated.

Dynamic Protein Chart

Dynamic Protein Binding

A 100 ml sample (3 mg/ml) of BSA was run on a 1 x 10 cm (8.0 ml) column at the indicated flow rates. The buffer was 50 mM Tris-HCl, pH 8.3. Elution was with 1.0 M NaCl.

Linear Flow Rate

Data for Macro-Prep High Q support was generated on a 14 cm (ID) X 17.4 cm (H) Amicon Moduline glass column with 1/4" plumbing. A Cole-Parmer Masterflex pump, size 18 neoprene tubing, Easy-Load pumphead was used. The eluent was water.

Common Buffers

Common Buffers for Ion Exchange

Type of Ion Exchanger	Buffer	Buffering Range
Cation	Acetic acid	4.8–5.2
	Citric acid	4.2–5.2
	HEPES	6.8–8.2
	Lactic acid	3.6–4.3
	MES	5.5–6.7
	MOPS	6.5–7.9
	Phosphate	6.7–7.6
	PIPES	6.1–7.5
	TES	7.2–7.8
	Tricine	7.8–8.9
Anion	Bicine	7.6–9.0
	Bis-Tris	5.8–7.2
	Diethanolamine	8.4–8.8
	Diethylamine	9.5–11.5
	L-histidine	5.5–6.0
	Imidazole	6.6–7.1
	Pyridine	4.9–5.6
	Tricine	7.4–8.8
	Triethanolamine	7.3–8.3
	Tris	7.5–8.0

Note:Buffer solution of 50 mM is recommended.

Macro-Prep 25 Q strong anion exchange support offers high-resolution separations at high flow rates with medium pressures. It possesses the same rigid, macroporous,and hydrophilic properties of 50 μm Macro-Prep High Q support, but in a 25 μm bead that offers higher resolution. Benefits include:

Benchtop to pilot-scale separations, where maximum sample recovery and purity are important
Fewer required chromatographic steps
Removal of protein variants and other difficult contaminants that would require additional validation if present in the final formulation