Affi-Prep Protein A Media

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Overview

Affinity Chromatography Resin Selection Guide
  Matrix Functional Group Specificity Capacity Working pH Pressure Limit Applications
Ready-to-Use Affinity Media
Nuvia™ IMAC High capacity, pressure-stable polymer based on UNOsphere beads NTA charged with Ni2+ Histidine ≥40 mg/ml* 2–14 45 psi
(3.1 bar)
Purification of recombinant histidine-tagged proteins; can be charged with other transition metals
Profinity™ IMAC Pressure-stable polymer based on UNOsphere beads IDA, available charged with Ni2+ and uncharged Histidine ≥15 mg/ml* 1–14 100 psi
(6.8 bar)
Suitable for purification of larger and difficult-to-purify recombinant histidine-tagged proteins; can be charged with other transition metals
Profinity GST** Pressure-stable polymer based on UNOsphere beads Immobilized glutathione GST-tagged proteins ≥10 mg/ml 1–14 45 psi
(3.1 bar)
Purification of recombinant GST-tagged proteins
Profinity eXact*** Crosslinked 6% agarose Subtilisin protease Subtilisin prodomain ≥3 mg/ml tag-free protein 2–10 10 psi (minus system pressure) Generation of native, tag-free proteins by on-column purification and cleavage
Affi-Gel® Protein A Crosslinked agarose Protein A
2 mg/ml
IgG   2–10 15 psi
(1 bar)
Purification of IgG from ascites, serum, and culture fluid; with MAPS buffer system, purification of 10 mg mouse IgG per ml of gel is possible
Affi-Prep® Protein A Pressure-stable polymer Protein A
2 mg/ml
IgG   2–10 1,000 psi (70 bar) Purified IgG from ascites, serum, and culture fluid; pressure-stable support for process-scale applications
Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA 1.9 mg/ml Albumin; general ≥11 mg/ml 2–10 15 psi
(1 bar)
Binds many nucleotide-requiring enzymes, albumin, and other proteins
DEAE Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA and DEAE Albumin and serum proteins 0.14 ml serum/ml gel 2–10 15 psi
(1 bar)
Purifies protease-free IgG from ascites, serum, and culture fluid with minimal sample preparation
CM Affi-Gel Blue Crosslinked agarose Cibacron Blue F3GA and CM Albumin and serum proteins 0.17–0.5 ml serum/ml gel 2–11 15 psi
(1 bar)
Produces albumin- and protease-free antibody preparation from serum without prior dialysis
Affi-Prep Polymyxin Pressure-stable polymer Polymyxin 2–4 mg/ml Endotoxins >5 mg/ml 2–10 1,000 psi
(70 bar)
Endotoxin removal
Affi-Gel Boronate Polyacrylamide gel Boronate 1.05 ± 0.15 cis-diols

meq/g
130 µmol sorbitol/ml 2–10 15 psi
(1 bar)
Adsorption of cis-hydroxyl–containing molecules, including sugars, nucleotides, and glycopeptides
Activated Media for Spontaneous Ligand Immobilization
Profinity Epoxide Pressure-stable polymer based on UNOsphere beads Epoxy group Nucleophiles; amini, thiol, -COOH 36–40 mg/ml lgG 1–14 Up to 80 psi
(5.5 bar)
Activated matrix for the immobilization of various ligands (for example, protein A, StrepTactin, and immunoglobulins)
Affi-Gel 10 Crosslinked agarose N-hydroxy-succinimide ≥10 µmol/ml -NH2 35 mg/ml 3–11 15 psi
(1 bar)
For coupling proteins with pI 6.5–11
Affi-Gel 15 Crosslinked agarose N-hydroxy-succinimide ≥9 µmol/ml -NH2 35 mg/ml 3–11 15 psi
(1 bar)
For coupling proteins with pI 6.5–11
Affi-Gel Hz Crosslinked agarose Hydrazide Oxidized carbohydrates 1–5 mg/ml 2–10 15 psi
(1 bar)
Immobilization of immunoglobulins and other glycoproteins via carbohydrate molecules
Affinity Media Using Carbodiimide Activation
Affi-Gel 102 Crosslinked agarose -NH2 16 ± 4 meq/ml -COOH 40 mg 2–11 15 psi
(1 bar)
Carbodiimide coupling of carboxyl-containing ligand

* Refer to bulletin 3193 for purification conditions.
** For example, Profinity GST resin is available only in prepacked cartridges.
***Profinity eXact Purification Resin.

Capacities of Protein A Supports
Immunoglobulin Affi-Gel Protein A
Capacity (mg/ml)
Affi-Prep Protein A
Capacity (mg/ml)
Mouse lgG1 6–10 7–9
Mouse lgG2a 6–10 7–9
Mouse lgG2b 6–10 7–9
Mouse lgG3 6–10 7–9
Mouse lgM* 6–10 7–9
Human lgG 15 10–12
Sheep lgG 8–10 7–9
Cow lgG 8–10 7–9
Horse lgG 8–10 7–9
Goat lgG 8–10 7–9
Rabbit lgG 8–10 7–9
Dog lgG 8–10 7–9
Pig lgG 8–10 7–9

* Approximately 50% of all mouse lgMs bind using the MAPS buffer system.

Binding Capacity Chart

Binding capacity for murine IgG subclasses. Mouse ascites fluids, containing equivalent amounts (5 mg) of either mouse IgG1, IgG2a, or IgG2b monoclonal antibody, were purified with 0.5 ml columns of Affi-Prep protein A matrix. Chromatography was performed with the Affi-Prep protein A MAPS II buffers.

Chromatography on Affi-Prep protein A media yields highly purified immunoglobulins (IgG), selectively removes IgG prior to analysis of other immunoglobulin classes, or adsorbs immune complexes for antigen purification. Protein A binds to the Fc region of immunoglobulins, especially IgGs from mammalian species.

The Affi-Prep protein A matrix allows high linear flow rates and exhibits excellent mechanical strength. These key features allow the direct transfer of experimental conditions from the initial analytical methods development stage to the final production scale purification of product. When the Affi-Prep protein A matrix is used in conjunction with the optimized Affi-Prep protein A MAPS II buffers, high efficiency binding of a wide range of immunoglobulins, including all isotypes of murine IgG, is achieved.

Advantages of the Affi-Prep protein A support include:

  • Binds a wide range of monoclonal and polyclonal antibodies, including mouse IgG1
  • Macroporous polymeric support is not compressible under normal operating conditions
  • Linear flow rates up to 2,000 cm/hr are achievable
  • High flow rates at all column diameters for easy scale-up
  • Compatible with most commonly used chromatographic buffers
  • Stable pH range from 2–10
  • Multiple chromatographic cycles without loss of capacity
  • High purity of IgGs
  • High affinity for mammalian IgG
  • High capacities for mouse IgG1 as well as other subclasses with MAPS optimized buffer
  • Easy to use
Affi-Prep Protein A Support

156-0006
5 ml, ready-to-use pressure-stable polymer support

List Price:   $389.00
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Affi-Prep Protein A Support

156-0005
25 ml, ready-to-use pressure-stable polymer support

List Price:   $1,848.00
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Number Description Options
5584 Bio-Scale Mini Cartridges, Product Information Sheet, Rev B Click to download
LIT230 Instruction Manual, Affi-Prep Protein A Matrix Click to download
LIT79 Instruction Manual, Affi-Prep Protein A MAPS II Kit Click to download
5712 The Profinia Protein Purification System Simplifies Antibody Purification With Protein A, Rev A Click to download [ Add to Cart (Free) ]
5726 Profinia Protein A and G methods Product Information Sheet, Rev A Click to download [ Add to Cart (Free) ]
5701 Profinia Buffers for Protein A and Protein G Chromatography PIS, Rev B Click to download [ Add to Cart (Free) ]