Introduction

CBinD^TM L is a new immobilized CBD-rProtein L matrix, designed for quick and efficient purification of immunoglobulins (Ig) and Ig fragments (Fab and scFv). Protein L binds primarily through the kappa light chain without interfering with the antigen binding site.

Applications

• Purification of IgG, IgA, IgE, IgD containing kappa light chains, especially human kappa light chains I, III and IV and mouse kappa light chain I
• Purification of scFv fragments containing kappa light chains as listed above
• Purification of human or mouse antibodies directly from cow, goat or sheep

Benefits

• Protein L from Peptostreptococcus magnus binds immunoglobulins (Ig) primarily through kappa light chain interactions without interfering with the antigen-binding site of Igs².
• Protein L binds to a wider range of Ig classes and subclasses from a wider variety of species than any other commercially available Ig binding protein.
• CBind^TML (CBD-rProtein L-cellulose) has improved binding capacity but similar binding properties than ProteinL-agarose.
• Very hydrophilic, and thus non-specific binding of proteins is minimal
• Designed to have enhanced flow properties

Properties

Properties	Description
Matrix:	Beaded cellulose
Ligand:	rCBDclos - Protein L
Coupling chemistry:	Bind & LockTM
Ligand density:	approx. 2.5 mg rCBD-Protein L /ml cellulose
Bead size range:	50-80 mm
pH stability	2.0 - 10
Storage buffer:	20% ethanol in PBS

Static Binding capacity for IgG:

Type	Value
Human:	approx. 17 mg/ml
Mouse:	approx. 16 mg/ml
Rat:	approx. 15 mg/ml
Rabbit:	approx. 2 mg/ml

Procedure for purification of antibodies (Ig)

Buffers:

• PBS: 20 mM K-phosphate buffer, 150 mM NaCl pH 7.2
• Elution buffer: 0.1 M Glycine pH 2.2
• Cleaning buffer: 6 M guanidine hydrochloride, 20 mM Tris/HCl pH 7.5

Sample preparation:

1. Clarify sample by centrifugation and/or filtration
2. Dilute or buffer exchange sample to equilibrate with PBS

Antibody purification:

1. Equilibrate CBindD^TML column with 5 column volumes of PBS.
2. Apply the sample. Effluent may be reloaded to improve purification yield.
3. Wash with 20 column volumes of PBS to remove the unbound material.
4. Elute purified antibodies with 2-5 column volumes of elution buffer.
5. If necessary the column can be cleaned with 6 M guanidine hydrochloride.
6. Re-equilibrate with 10 column volumes of PBS.
7. Neutralize eluted antibodies with 1 M Tris base.

This procedure has been successfully applied to the purification of Ig from human plasma, monoclonal IgG from mouse ascites fluid, single chain antibodies (ScFv) derived from E.coli and Fab fragments, prepared by partial digestion of human IgG with papain.

Product Range

Cat. No.	Product	Package Size
59898	rCBD-ProteinL	1 mg, 5 mg
34431	rCBD-ProteinL - Cellulose	1 ml, 5 ml suspension in PBS

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