Select a link below to access Protocols for Abundant Protein Removal.

• Abundant Protein Removal Using the Enchant™ Albumin Depletion Kit (2.1.2)
• Abundant Protein Removal Using the Enchant IgG Purification Kit (2.1.3)
• Abundant Protein Removal Using the Enchant Multi-Protein Affinity Separation Kit (2.1.4)
• Abundant Protein Removal Using BioSepra™ Chromatography Resins (2.1.5)

Introduction
Complexity reduction in samples is an important step in facilitating access to the low abundant proteins of interest for disease research and diagnostics. This process for human serum and plasma frequently includes the depletion of high abundant proteins such as albumin and IgG in combination with other fractionation techniques prior to 2D gel electrophoresis or LC-MS/MS detection.

Pall currently offers several solutions for abundant protein separation or removal. Options include affinity chromatography resins as well as complete kits for removal of multiple abundant proteins (albumin, IgG) in one step.

Enchant Protein Separation Kits rapidly deplete abundant proteins and unmask low abundant biomarkers from serum and plasma samples. Alternatively, these kits can be used to collect the abundant protein fractions for further analysis.

Enchant Protein Separation Kits

Abundant Protein Separation Quick Selection Guide

Choose the Best Protein Removal Product for Your Needs

 

Select a link below to access Protocols for Protein Fractionation (Complexity Reduction).

• Protein Fractionation (Complexity Reduction) Using Q, S, DEAE, and CM Ion Exchange Media (2.2.2)
  • Ceramic HyperD® Ion Exchange Resin (2.2.2.1)
  • Mustang® Ion Exchange Membrane (2.2.2.2)
• Affinity Separations — Using Chromatography Resins for Protein Fractionation (Complexity Reduction) (2.2.3)
  • Heparin HyperD M Resin (2.2.3.1)
  • Lysine HyperD Resin (2.2.3.2)
  • IMAC HyperCel™ Resin for Immobilized Metal Ion Affinity Chromatography (2.2.3.3)
  • Protein A Ceramic HyperD F Resin (2.2.3.4)
• Protein Fractionation (Complexity Reduction) Using Mixed Mode Chromatography Resins (2.2.4)
  • MEP HyperCel Mixed Mode Resin (2.2.4.1)
  • HA Ultrogel® Resin (2.2.4.2)
• Protein Fractionation (Complexity Reduction) Using Size Fractionation Chromatography Resins (2.2.5)
  • Ultrogel AcA Size Exclusion Chromatography Resin (2.2.5.1)

Introduction
Sample complexity reduction is an important first step to facilitating access to the low abundant proteins of interest for clinical research and diagnostics. The process for human serum and plasma frequently includes the depletion of highly abundant proteins such as albumin and IgG in combination with other fractionation technologies, such as ion exchange or affinity separation, prior to 2D gel or LC-MS/MS analyses.

Pall currently offers several solutions for small-scale chromatographic protein fractionation (see table below). Options include chromatography resin for affinity capture, ion exchange separation, and mixed-mode capture resins. Pall also offers membrane chromatography solutions, i.e., Mustang membranes. All chemistries are available for use in several forms including standard chromatography columns, disposable columns, spin filters, and multi-well filter plates.

Protein Fractionation Quick Selection Guide

Available Separation Columns

Description	Column Volume	Available from Pall
Glass Chromatography Column	Varies	No
Disposable Chromatography Column	Varies	No
Spin Filter	< 1 mL	Yes
Deep Well Multi-Well Filter Plate	96 x < 1 mL	Yes
Multi-Well Filter Plate	96 x < 350 µL	Yes

Select the link below to access a Protocol for Detergent Removal.

• Using SDR HyperD Solvent and Detergent Removal Chromatography Resin (2.3.1)

Select a link below to access Protocols for Concentration, Desalting, and Buffer Exchange.

• Concentration and Desalting Using Ultrafiltration (2.4.2)
  • Concentration and Diafiltration of Samples (< 60 mL) in a Spin Filter (2.4.2.1)
  • Desalting and Buffer Exchange of Samples (< 1 mL) in an AcroPrep™ 96 and 384 Multi-Well Filter Plate (2.4.2.2)
• Desalting with Size Exclusion Media (2.4.3)
  • Ultrogel AcA 202 and Trisacryl® GF-05M Resins (2.4.3.1)

Introduction
Biomolecule purification involves a complex series of steps where targets are selectively separated through sequential processes. The processes by which separation is performed often creates a need for the sample to be desalted or concentrated to prepare the biomolecule sample for the next step in the purification process.

Pall offers several technologies to perform efficient sample concentration, desalting, and buffer exchange including ultrafiltration (UF) spin filters, UF multi-well filtration plates, and bottled gel filtration media. Refer to the table below for a selection of UF products available.

UF Products for Concentration and Desalting

Device	Sample Volume
AcroPrep 384 filter plate	< 100 µL
AcroPrep 96 filter plate	< 350 µL
Nanosep® device	< 0.5 mL
Microsep™ device	0.5-3.5 mL
Macrosep® device	3-15 mL
Jumbosep™ device	15-60 mL

Compared to other methods, UF methods for concentration and desalting offer a number of advantages including:

• Reduction of processing steps — desalting and concentration take place in a single step.
• Consistency — the use of UF products eliminates problems with gel beds.
• Versatility — UF is useful in both low and high throughput modes.

Pall also offers chromatography resins for desalting applications including Ultrogel AcA 202 and Trisacryl GF-05 resins. Refer to the below table for general properties.

Properties of Ultrogel AcA and Trisacryl GF-05 Resin

Specification	Ultrogel AcA 202 Resin	Trisacryl GF-05M Resin
Particle Size	60-140 µm	40-80 µm
Monomer	20% (w/v) Acrylamide	N-acryloyl-2-amino-2-hydroxymethyl-1,3-propanediol
Cross-linker	2% (w/v) Agarose	Hydroxylated acrylic bifunctional monomer
Exclusion Limit	22,000 dalton	3,000 dalton
Linear Fractionation Range	1,000-15,000 dalton	200-2,500 dalton
Resolving Power (plates/m)	3,000	2,500
Working pH Range	3-10	1-11

Pall Centrifugal Spin Filters

Select a link below to access Protocols for Generic Clarification of Samples by Microporous Filtration (Particulate Removal).

• Clarification of 0.1-1.0 mL Samples Using AcroPrep 96 and 384 Multi-Well Filter Plates (2.5.2)
• Clarification of Samples (< 1 mL) in Spin Filters (Nanosep® Centrifugal Devices) (2.5.3)
• Clarification of Samples (1-100 mL) in an Acrodisc® Syringe Filter (2.5.4)
• Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration Device (2.5.5)
• Clarification of Plasma or Serum Samples to Remove Cryoprecipitate Using Acrodisc Syringe Filters with a Prefilter (2.5.6)
• Prefiltration of Plasma or Serum Samples Using Acrodisc Syringe Filters with Glass Fiber (2.5.7)

Introduction
Although a basic filtration concept, the clarification and prefiltration of samples remains an important function within protein sample preparation applications. When filtration is used as a prefilter, matching the proper filter media and device to the application is critical. Large pore size filter materials are used to filter solutions prior to more detailed analysis. When selecting the best product for your application, several factors need to be considered. Sample viscosity, sample volume, and sample recovery are just some of the aspects that will drive the selection of the optimal device.

Pall offers a number of media and device options for fast, effective filtration with minimal sample hold-up for both single and high throughput sample processing. From sample volumes of a few microliters to multiple liters, Pall can supply the best product solution for your application.

Choose the Best Clarification Device for Your Needs

Acrodisc Syringe Filters for Analytical Sample Preparation

Select the link below to access a Protocol for Endotoxin Removal.

• Endotoxin Removal from Water Buffer, Neutral Sugars, and Certain Biological Solutions (2.6.1)