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Immunoaffinity fractionation of highly-abundant proteins (HAP) has proven to be one of the most effective approaches for overcoming the wide dynamic range of protein concentration, as well as enabling the detection of low abundant proteins (LAP). Avian polyclonal IgY (Immunoglobulin Yolk) antibodies have unique and advantageous features that allow for highly-specific fractionation of a complex protein mixture. The IgY14 column can effectively remove 14 HAP from plasma. The next level of abundant proteins, moderately abundant proteins (MAP), becomes an obstacle to access LAP, where the majority of biologically interesting and clinically important biomarkers reside. Therefore, isolation of MAP is a new challenge for effectively and accurately detecting and analyzing LAP.
To tackle this challenge the SuperMix column was developed to separate MAP from LAP. The SuperMix System was developed by immunizing chickens with a flow-through fraction of IgY-12 or IgY14 column and constructing the column with affinity-purified IgY antibodies against the flow-through proteins of IgY-12 or IgY14.
SuperMix Fractionation Process
The Human SuperMix column is then used in conjunction with the IgY14 column to provide superior depletion.
SuperMix Results
As shown below, the SuperMix column was applied for fractionating the flow-through fraction of IgY14, which resulted in a bound/eluted fraction (designated MAP fraction) and the flow-through fraction (designated LAP fraction). Unfractionated and serial-fractionated samples using IgY14 and SuperMix columns were analyzed by SDS-PAGE and 2DE. Our data demonstrate that SuperMix columns specifically and reproducibly remove a group of proteins, which are the main abundant proteins in the flow-through fraction of IgY14.
 
| Product Name |
Column Use |
Capacity |
Product # |
SuperMix LC-2 Column Kit
Includes single SuperMix LC-2 column and reagents. |
100 times |
Flow-through from IgY14 LC-5 column |
SEP050 |
SuperMix LC-5 Column Kit
Includes single SuperMix LC-5 column and reagents. |
100 times |
Flow-through from IgY14 LC-10 column |
SEP060 |
Recommended Set-Up:
| Product Name |
Column Use |
Human or Primate plasma |
IgY14 LC-10/SuperMix LC-5
Columns are run in conjunction for intensive mass spectrometry and 2D-gel analysis |
100 times |
250µl |
IgY14 LC-5/Supermix LC-2
Columns are run in conjunction for intensive mass spectrometry and 2D-gel analysis |
100 times |
125µl |
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