Macro-Prep 25 S Cation Exchange Media

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Overview

	Macro-Prep^® 25 Q Support	Macro-Prep 25 S Support
Type of ion exchanger	Strong anion	Strong cation
Functional group	–N⁺(CH₃)₃	–SO₃^-
Ionic capacity (µeq/ml)	220 ±40	110 ±30
Typical binding capacity	>30 mg/ml BSA	>40 mg/ml IgG
Shipping counterion	Cl^-	Na⁺
Nominal particle size	25 µm	25 µm
Nominal pore size	725 Å	725 Å
Max linear flow rate	3,000 cm/hr	3,000 cm/hr
Chemical stability: 1% SDS, 24hr 6M guanidine-HCI, 24 hr	Yes Yes	Yes Yes
Volume changes: pH 4–10 0.1–1.0 M NaCl	<1% <5%	<1% <4%
Autoclavable (121°C, 30 min)	Yes	Yes
pH stability	1–10	1–10
Antimicrobial agent	20% ethanol	20% ethanol

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.

Common Buffers

Common Buffers for Ion Exchange

Type of Ion Exchanger	Buffer	Buffering Range
Macro-Prep 25 S	Acetic acid	4.8–5.2
	Citric acid	4.2–5.2
	HEPES	7.6–8.2
	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
	Pivalic acid	4.7–5.4
	TES	7.2–7.8
	Tricine	7.8–8.9
Macro-Prep 25 Q	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

Linear Flow Rate

Linear Flow Rate vs. Operating Pressure

The Macro-Prep 25 S support was evaluated for linear flow rate vs. operating pressure by measuring the pressure at a series of flow rates using 1M NaCl on a 1.6 cm (ID) x 12.5 cm (H) column. The pressure plotted was determined by subtracting the pressure generated by the column when empty from the pressure generated from the column when packed.

Separation on Macro-Prep 25 S

Separation of a mixture of 1 mg of native protein from 0.23 mg of the protein with oxidized surface methione residues from treatment with t-butyl hydroperoxide on a 1.6 cm (ID) x 12.5 cm (L) column packed with Macro-Prep 25 S.Buffer A: 50 mM KH₂PO_4, pH 5.0. Buffer B: 50mM KH₂PO_4, 2 M KCl, pH 5.5; gradient: 50% B to 100 % B in 20 min; flow rate: 2 ml/min.

Macro-Prep 25 S strong cation exchange support offers high-resolution separations at high flow rates with medium pressures. It possessesthe same rigid, macroporous, and hydrophilic properties of 50 μm Macro-Prep High Q and High S media, 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