Depletion Column

An essential tool for proteomic study and biomarker discovery

Why the depletion columns are required.

In order to discover and identify disease-associated biomarkers, the proteomics technology platform most commonly used consists of high resolution protein separation by two-dimensional electrophoresis (2-DE), mass spectrometric identification of proteins from stained gel spots and a bioinformatic data analysis process supported by statistics. Profiling proteins and their disease- or treatment-related change is much more easily achieved in tissue homogenates than in plasma samples. Plasma protein display and quantitation suffer several disadvantages including very high abundance of certain proteins; high heterogeneity of many proteins resulting in long charge trains; crowding of 2-DE separated protein spots in the molecular mass range between 45-80 kDa and in the isoelectric point range between 4.5 and 6 (1). A prefractionation step, such as depletion of a few high-abundant proteins before protein profiling, can assist in the discovery and detection of less abundant proteins that may prove to be informative biomarkers (2).
 
Reference: 
1.Pieper R, Su Q, Gatlin CL, Huang ST, Anderson NL, Steiner S. Multi-component immunoaffinity subtraction chromatography: an innovative step towards a comprehensive survey of the human plasma proteome. Proteomics. 2003; 3(4):422-32.
2. Björhall K, Miliotis T, Davidsson P. Comparison of different depletion strategies for improved resolution in proteomic analysis of human serum samples. Proteomics. 2005; 5(1):307-17. 
 

Why the IgY(ΔFc)-based affinity column is your preferred choice.

1) Unique Characteristics
Due to a largely phylogenetic difference between duck and the antigens of mammalian, IgY(ΔFc) can recognize more epitopes and is relatively more reactive to the highly conserved protein than mammalian IgG. Moreover, IgY(ΔFc) do not bind some plasma/serum proteins (such as complement factors, rheumatoid factor etc.), thus avoiding the risk of losing these proteins.
 
2) Specific Binding
The major drawback in albumin depletion by using the blue-dye-based mediums is lacking selectivity. In addition to albumin, the blue-dye also adsorb more than 50 other plasma/serum proteins including Alpha-2-marcoglobulin、Ceruloplasmin、Alpha-1-antitrypsin、apolipoprotein A-I、antithrombin-III、C1 inhibitor、complement factor B、kininogen、angiotensin、gelsolin、plasminogen and heparin cofactor II etc. Consequently, these proteins will be lost during the depletion process. Moreover, protein A resin does not fully bind all subclasses of human IgG such as IgG3, plasma/serum proteins or informative biomarkers is greatly minimized. resulting in incomplete depletion. The GBC Human Albumin/IgG/Transferrin 3 in 1 Depletion Column is designed to specifically remove albumin, IgG and transferrin from human serum/plasma. By using GBC Human Albumin/IgG/Transferrin 3 in 1 Depletion Column, the risk of losing other plasma/serum proteins or informative biomarkers is greatly minimized.
 
3) High depletion rate
The GBC 3 in 1depletion column can efficiently remove albumin, IgG and transferrin proteins from plasma/serum, which removes greater than 98% of the albumin, IgG and transferrin.
 

GBC Human Albumin/IgG/Transferrin 3 in 1 Depletion Column/Kit feature in:

Efficient removal of albumin, IgG and transferrin with minimal loss of other serum biomarkers
Fast and simple procedure, applicable for multiple processing
Not bind mammalian complement factors, rheumatoid factor, IgM, Fc receptor
Re-usable
Applications: 1D or 2D electrophoresis, LC/MS, etc.
 
Product name
 
(REF)
GBC 3 in 1 Depletion
Spin Column
GBC 3 in 1 Depletion
Spin Column Kit
GBC 3 in 1
Depletion Column
Depletion Method
Spin-down
Spin-down
Gravity-flow
Column Size
1.5 mL
1.5 mL
10 mL
Resin Beds
0.32 mL
0.32 mL
4 mL
Sample Loading volume
15 µl
15 µl
200 µl
Depletion rate:
Albumin
IgG
Transferrin
>98%
>98%
>98%
Enriched protein
(per column/per run)
280-330 µg
280-330 µg
3.0-3.4 mg
Cycle for use*
more than 20 cycles
more than 20 cycles
more than 200cycles
Package
Spin Column x 2
Spin Column x 2
Equilibration Buffer 80mL
Elution Buffer 60mL
Depletion Column x 1
*The numbers of cycle for use are dependent on the individual operation.