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Protein L Agarose Beads / Resin

DatasheetSpecific ReferencesReviewsRelated ProductsProtocols
 
Introduction
 

Sino Biological rProtein L resin was produced by immobilizing recombinant protein L to 4% cross-linked agarose matrix. It is designed for easy, one-step purification of classes, subclasses and fragments of immunoglobulins.

 
Specifications
 
Ligand Recombinant protein L
Binding capacity 10-15 mg human IgG/ml
Matrix Spherical, 4%  agarose
Average particle size 90μm(45-165μm)
Maximum linear flow >150 cm/h
Maximum Pressure 0.3 MPa
pH stability pH 3-9
Storage 20% ethanol at 2°C to 8°C
 
Experiment
 
Column: 5x 50 mm, bed volume 1.0 ml
Resin: Protein L 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 binding 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 column with binding buffer, when asorbency of 280 nm decrease to base line, elute with elution buffer.
Result: 16 mg antibody was recovered in eluant, Dynamic binding capacity of the column is about 16 mg/ml.

 

 

Chromatography graph of measuring dynamic binding capacity

 
Protein L 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 L Resin: CIP Protocol
 

Wash the column with 8 M urea, 0.05 M citric acid pH 2.5. Immediately re-equilibrate with at least 5 bed volumes of binding buffer.

 
Protein L 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 L 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.

 
IgG 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 G resin
Recombinant Protein A
Recombinant Protein G
Recombinant Protein L

 

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Catalog: 11044-H07E-RN-25
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