Host Cell Protein (HCP) by DIGE vs. ELISA

Advantages of DIGE Western Blot Over ELISA and Standard 2D In HCP Detection

	ELISA	Standard 2D Gel	2D DIGE
Number of gels for each Ab		Two	One
Detection Method		Silver Staining	CyDye Labeling
HCP Composition and profiling	No
Detect protein modification	No
Detect protein degradation	No
Protein and WB in same gel	No	No
In-gel protein and WB comparison	No	No
Accuracy	Low	Low	High
	Inaccurate : In-direct reaction of antibodies with the antigen’s associated proteins	No QC or Detection of transferred proteins on Membrane	Protein image is scanned on the membrane
	False negative: Coated Ab does not capture all HCP; Surface denaturation of coating Ab can affect detection	Error in matching spots between gel to gel, and gel to the transferred membrane	Protein and Antibody signal on the same gel, thus avoiding any ambiguity
Consistency and Reproducibility	Lower	Lower	Higher