Spin column-based Antibody Purification Kit 

Monolithic silica column

Monolithic silica column for antibody purification

Spin	Protein A
	Protein G

 Performance of column with a commercially available antibody drugs

Product specification	Carrier	Monolithic silica gel
	Binding capacity （Human IgG）	Spin	0.4mg/column
	Maximum amount of sample	Spin	0.5mL
Kit	Column・Sampling tube Various solutions（Adsorption, washing, elution, reproduction, and neutralizing）

Spin-type

Antibody purification in a short time becomes possible by immobilizing the Protein A and Protein G on the plate-shaped monolithic silica surface.　Purification and concentration of target IgG antibody, and measurement of IgG antibody concentrations in the　culture solution can be performed easily in two minutes.　*The high-density chemical bonding technology into a modified protein G and the silica surface. Using this technique, monolithic silica column for antibody purification.　Before using these products, please thoroughly read and understand instructions stated in instruction manual.

 

Performance 

Protein Ａ（ProA)	Protein Ｇ（ProG）
Up to 0.4mg of Human IgG can be purified at a recovery rate of 90% or more by using a spin column ligand with Ex-Pure ProA.　In addition, up to 0.3mg of protein can be quantitatively measured in each step.	A spin column ligand with Ex-Pure ProG can purify and recover antibodies as easily as Ex-Pure ProA, and the amount of protein within the liquid passage can be quantitatively measure in each step. Furthermore, IgG3 type antibodies, which cannot bind to Ex-Pure ProA, can also be purified and recovered.

 Evaluation of antibody recovery accuracy (variation) of the spin column

Operating procedures

1 . Prepare 10 spin columns ( ProA or ProG ) at random .
2 . IgG from human serum (11.7mg/ml) from Sigma was used to increase and adjust the IgG concentration to 2.0 mg/ml.
3. Apply 0.4ml of the IgG sample, and perform the following steps: washing → elution 1 → elution 2. Elution 1, Elution 2
　 The amount of recovered IgG antibody was quantified by the UV method (A280) and by BCA.

UV method	Protein Ａ（ProA)	Protein Ｇ（ProG）
	0.65±0.027mg/column	0.504±0.056mg/column
BCA method	0.641±0.079mg/column
0.4mL of 2.0mg/mL human IgG was injected into a randomly selected spin column, and steps from washing to elution were performed. 　The antibody content in the elution solution was measured using the UV method (A280) and the BCA method. *For Ex-Pure ProG, only the UV method was used.

Protein A - Amount of antibody added and recovered

Protein G - Amount of antibody added and recovered

 Protein G - IgG3 antibody added and recovered

 Protein A - Human IgG antibody added and recovered

 

Antibody Recovery - Commercially available monoclonal IgG antibodies
(Rituxan, Avastin, Herceptin)