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Protein G Agarose Beads / Resin

DatasheetSpecific ReferencesReviewsRelated ProductsProtocols
 
Introduction

Sino Biological Protein G Agarose Beads / Resin was produced by immobilizing recombinant protein G to 4% cross-linked agarose matrix. It is designed for easy, one-step purification of classes, subclasses and fragments of immunoglobulins or recombinant proteins containing Fc regions from biological fluids or cell culture media.

 

 
Specifications
 
Ligand Recombinant Streptococcal Protein G
Dynamic binding capacity ~20 mg/ml
Matrix Spherical, 4% agarose
Average particle size 90um(45-165um)
Maximum linear flow rate >150 cm/h
Maximum Pressure: 0.3 MPa
pH stability  Long term:
Short term:
pH 3-9
pH 2-10
Storage 20% ethanol at 2°C to 8°C
 
Experiment:
Column: 5x 50 mm, bed volume 1.0 ml
Resin: Protein G agarose resin
Binding Buffer: 50 mM Tris, 100 mM NaCl, pH 8.0
Elution Buffer: 100 mM glycin, 10 mM NaCl, pH 3.0
Sample: humanized monoclonal antibody,1.9 mg/ml
Flow Rate: 0.3 ml/min
Procedure: The column was equilibrated with biding buffer, then apply the sample at fow rate of 0.3 ml/min, when the flowthrough concentration reach 0.19 mg/min stop loading and wash the columm with biding buffer, when asorbency of 280 nm decrease to base line, elute with elution buffer.
Result: 20 mg antibody was recovered in eluant, Dynamic binding capacity of the colun is about 20 mg/ml.

 

Chromatography graph of measuring dynamic binding capacity

 
Protein G Resin: Repeated Cycle Usage

The resin is reusable with proper regeneration procedure, no significant recovery loss or performance change after 20 cycle usage.

SDS-PAGE of monoclonal antibody purified after 20 repeat usage cycle

 
Protein G Resin: CIP Protocol

Remove strongly bound hydrophobic proteins, lipoproteins and lipids by washing the column with a non-ionic detergent (e.g. Triton™ X-100), 0.1%, at 37°C, Immediately re-equilibrate with at least 5 bed volumes of sterile filtered binding buffer. Alternatively, wash the column with 70% ethanol and let stand for 12 hours. Re-equilibrate with at least 5 bed volumes of sterile binding buffer.

 
Protein G Resin: Stability Study

The IgG binding capacity and recovery was maintained after seven days storage at room temperature in solutions list below:
3 M NaSCN
6 M guanidine-HCl
8 M urea
1 M acetic acid, pH 2.0
0.1 M glycine, pH 3.0
70% ethanol.

 

Recovery of antibody at the loading burden of 18 mg/ml medium after storage in varous solutions

 
Protein G Resin: Column Regeneration

Regenerate column by washing with 3-5 bed volumes of Elution Buffer and then re-equilibration with Binding Buffer. In some applications, substances like denatured proteins or lipids, do not elute in the regeneration procedure. These can be removed by cleaning-in-place procedures.

 
Protein G Resin: Storage

For storage, keep the medium at 2°C to 8°C in a suitable bacteriostat, e.g. 20% ethanol and/or 0.02% sodium azide. Notice: The resin must not be frozen.

 

 
gG binding of protein A, G, and L

 

Species

Protein A

Protein G

Protein L

 Human

 IgG

+++

+++

+++

 

 IgG1

++++

++++

++++

 

 IgG2

++++

++++

++++

 

 IgG3

-

+++

+++

 

 IgG4

++++

++++

++++

 

 IgA

+

-

+++

 

 IgA1

+

-

+++

 

 IgA2

+

-

+++

 

 IgD

-

-

+++

 

 IgE

++

-

+++

 

 IgM

+

-

+++

 Rabbit

 IgG

+++

+++

+

 Cow

 IgG

+

+++

-

 

 IgG1

+

+++

-

 

 IgG2

+++

+++

-

 Cat

 IgG

+++

+

?

 Horse    

 IgG

++

++++

?

 Goat

 IgG

+

++

-

 

 IgG1

+

+++

-

 

 IgG2

+++

+++

-

 Guinea-pig

 IgG1

++

+

?

 

 IgG2

++

+

?

 Sheep

 IgG

+

++

-

 

 IgG1

+

++

-

 

 IgG2

+++

+++

-

 Dog

 

++

+

?

 Pig

 

+++

++

+++

 Rat

 IgG

+

++

+++

 

 IgG1

-

+

+++

 

 IgG2a

-

++++

+++

 

 IgG2b

-

++

+++

 

 IgG2c

++

++

+++

 

 IgG3

+

++

?

 Mouse

 IgG

++

++

+++

 

 IgG1

+

++++

+++

 

 IgG2a

++++

++++

+++

 

 IgG2b

+++

+++

+++

 

 IgG3

++

+++

+++

 

 IgM

-

-

+++

 Chicken

 IgY

-

-

-

 Monkey(rhesus)

 IgG

++++

++++

?

 Hamster

 

+

++

+++

 Koala

 

-

+

?

 Llama

 

-

+

?

 

Strong binding ++, medium interaction +, weak or no interaction -.

 
Related products

 

Ultra-linked rProtein A resin
rProtein L resin
Recombinant Protein A
Recombinant Protein G
Recombinant Protein L

 

 

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