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IgG Purification Tool Beyond Protein A and G

Purification of IgG - Affinity Gel 【Ab-Capcher™】, Column 【Ab-Rapid PuRe™】, Spin Column 【Ab-Rapid SPiN™】

Background

Ab-Capcher is "Protein A-R28" (Protein A variant) which is developed with ProteNova’s patent technology bound at high density to a supporting gel matrix offering many practical advantages for routine purification of rat and mouse IgG including broader subclass specificity and high capacity binding.

Ab-Rapid PuRE is a specially designed syringe-type column unit prepacked with AbCapcher offering maximum convenience for Ig purification.

Ab-Rapid SPiN is a convenient spin column prepacked with Ab-Capcher.

 

Protein A-R28 is an alkali-tolerant IgG-binding protein derived from protein A, which is developed with ProteNova’s patent technology. Protein A-R28 strongly binds to various species and subclasses of IgG, compared with Protein A and G. The coupling to resin (Ab-Capcher) provides an alkali-washable unique affinity medium with high binding capacity for immunoglobulin, which is useful for purification of human, rabbit, and mouse IgGs including mouse IgG1.Ab-Aapcher is also useful for immuno-precipitation experiments.

Feature and Advantages

 5 times the binding capacity of Protein G
 Labile antibodies can be bound at neutral pH
 Low non-specific binding
 IgG puirty >95%

Binding Characteristics of Ab-Capcher
Species SubclassAb-CapProtein AProtein G
Mouse IgG1 ++++ + ++
IgG2a +++++ ++++ +
Rat IgG1 +++ - +
IgG2a -* - +
Goat IgGs ++++ - +
Chicken IgY - - -
Human IgG +++++ ++++ ++
Rabbit IgG +++++ ++++ ++

*Connection has not confirmed.

Characteristic

Composition : Highly cross-linked 4% agarose
Particle size : 45-165 um
Ligand : Alkali-+tolerant variant of Protein A (Protein A-R28) (E.coli)
Binding capacity (max.) : > 65mg human IgG / ml gel
Delivery conditions : 20% EtOH

Recommended conditions

< Example of suitable buffers for human IgG (-70 mg / ml gel) >
Binding buffer : 20 mM sodium phosphate buffer, 150 mM NaCl, pH 7.2
Elution buffer : 0.1 M Glycine-HCL, pH 2.8
Neutralization buffer : 1 M Tris ( add 1/20 vol of elution buffer)
Washing buffer : 0.1 N NaOH
< Example of suitable buffers for mouse IgG1 (-40 mg / ml gel) >
Binding buffer : 1.5 M Glycine-HCL, 3 M NaCl, pH 9.0
Elution buffer : 0.1 M Glycine-HCL, pH 2.8
Neutralization buffer : 1 M Tris ( add 1/20 vol of elution buffer)
Washing buffer : 0.1 N NaOH
# Phosphate buffered saline can be used as binding buffer
(in this case, binding capacity is approx 20 mg of mouse IgG1 / ml of the gel)

Application DATA

Binding of Human IgG
Binding capacities of Ab-Capcher and two protein A-resins were tested with human gamma globulin. Phosphate buffered saline (PBS) was used as the binding buffer. Ab-Capcher showed the highest binding capacity in affinity media tested (-70 mg human IgG / ml gel).

Binding of monoclonal mouse IgG1 at pH 9
Mouse ascites were precipitated with 50%-saturated
ammonium sulfate, dissolved and dialyzed with PBS. The fraction diluted 3-fold with 1.5 M Glycine / 3 M NaCl buffer (pH 9.0). was used to test binding capacity of Ab-Capcher For mouse IgG1. Ab-Capcher bound high amounts of mouse IgG1 at pH 9.0 as well as human IgG pH 7.0.

Binding of monoclonal mouse IgG1 with PBS
Fraction of 50%-saturated ammonium sulfate from mouse ascites was used to test binding capacity of Ab-Capcher and Protein G-agarose for mouse IgG1 under physiological conditions. PBS was used as binding buffer. Ab-Capcher bound larger amounts of mouse IgG1 about 5-times than conventional Protein G-agarose under physiplosical conditions ( 18.3 mg / ml gel ).

Alkali-arability
Relative binding capacities for human IgG of Ab-Capcher and conventional Protein A-agarose were measured after treatment with 0.1 N NaOH at 25℃. Ab-Caocher remains 90 % of binding capacity after treatment for 106 hr at 25℃.

 

< Experiment example data > ・・・ coming soon

<Ab-Capcher FAQ> ・・・ coming soon

Product List

Product Name Cat# Quantity Price

Ab-Capcher

PTN-P-002-2 2ML

¥ 25,000
$ 334
€ 250

Ab-Capcher

PTN-P-002-10 10ML

¥ 75,000
$ 1000
€ 750

Ab-Rapid Pure 2

PTN-P-012-2 2COLUMN

¥ 16,000
$ 214
€ 160

Ab-Rapid Pure 10

PTN-P-012-10 10COLUMN

¥ 64,000
$ 854
€ 640

Ab-Rapid SPiN10

PTN-P-013-10 10PC

¥ 20,800
$ 278
€ 208

Ab-Rapid SPiN50

PTN-P-013-50 50PC

¥ 68,400
$ 912
€ 684

Buffer kit for Ab-Rapid Pure

PTN-P-011 10RXN

¥ 5,000
$ 67
€ 50

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.