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> Tech Support > Host Cell Protein (HCP) by DIGE vs. Elisa

HCP Analysis: Advantages of 2D DIGE Western Blot Over ELISA & Standard 2D WB




  ELISA Standard 2D Western Blot 2D DIGE Western Blot

Number of gels for each Ab

  Two One

Detection Method

  Silver Staining CyDye Labeling

HCP Composition and profiling


Detect protein modification


Detect protein degradation


Protein and WB in same gel

No No

In-gel protein and WB comparison

No No


Low Low High

Inaccurate: In-direct reaction of antibodies with the antigen’s associated proteins

Protein spots are counted from the gel, NOT from the membrane

Transferred protein spots are scanned directly from the membrane

False negative: 1) Coated Ab does not capture all HCP; 2) Surface denaturation of coating Ab can affect detection

Error caused by: 1) Gel to gel variation; 2) Gel to membrane variation

Accurate: Protein and Antibody signal on the same gel, thus avoiding any ambiguity

Consistency and Reproducibility

Lower Lower Higher