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Serum Biomarker Discovery By 2D DIGE Proteomics Analysis




In serum and plasma, the expression leves of low and high abundant proteins exceeds 10 orders of magnitude and this wide dynamic range prevents the discovery of low abundant disease-specific serum markers. Immunodepletion has been used to enhance the detection of low abundance plasma proteins. However, researches have discovered that, even with immunodepletion, low abundance proteins still remain undetectable using LCMSMS platform. Furthermore, immunodepletion process may remove proteins associated with the abudant proteins and cause protein degradations, thus creating more complexities.

Applied Biomics developed a technology by which thousands of low-abundance serum proteins can be resolved and quantitatively analyzed on 2-D gels without immunoaffinity depletion of the serum abundant proteins. This technology allows for a more sensitive detection and accurate quantification of low-abundance plasma proteins. For details please view see our Publication on Genetic Engineering & Technology News. Such platform has been successfully applied in discovering disease- or treatment-specific serum biomarkers.

* Example 1: Human Serum Sample.
The left, middle and right image shows the human serum proteome using standard protocol, Applied Biomics (ABO) improved protocol with pH3-10, and pH4-7, respectively. The ABO protocol clearly reveals much more low aboundant serum proteins than the standard protocol.

Standard 2D Protocol (pH3-10)

Serum 2D DIGE
ABO pH3-10 Protocol

 Serum 2D DIGE
ABO pH4-7 Protocol

 Serum 2D DIGE


* Example 2: Preclinical Trial Mouse Serum Before and After Drug Feeding Using ABO Protocol. Four pairs of mouse serum samples were collected before and after 4 different treatments, then analyzed by using Applied Bimics' improved serum protocol. The overlay images showed the difference between the serum samples collected before (green color) and after (red color) the drug treatment. The circled spots (in red) indicated a protein with increased abundance due to drug treatment. The 3-D view of the same spot is displayed in the right panel.

Serum 2D DIGE

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