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Host Cell Protein (HCP) Assay: Antibody Coverage
Host Cell Protein (HCP) Assay: Product Purity

 

 

 

Click to view a complete list of Applied Biomics HCP assays

Applied Biomics Host Cell Protein (HCP) assays using 2D DIGE followed by fluorescent Western blot offer significant ADVANTAGES over the ELISA and standard 2D Western blot, with the following applications:

1) Characterizing HCP antibody coverage
2) Detecting HCP contaminants originated from cells that are used to manufacture therapeutic proteins and vaccines

Experimental Procedure: HCP 2D Western Blot (HEK293)
Example-1: CHO Cell HCP Assay
Example-2: E.coli HCP Assay
Example-3: E.coli High Mw HCP Assay
Example-4: Yeast (Saccharomyces cerevisiae) HCP Assay
Example-5: Characterization of SF9 HCP antibodies from same 2D gel

Advantages of DIGE over ELISA and Standard 2D in HCP Detection

Applied Biomics will perform HCP assays using HCP antibodies provided by customers. In addition, we cover the following HCP antibodies:

Bacteria Mammalian Cell Yeast Insect Others

E.coli HCP
Lactococcus lactis HCP





Vero Cell HCP
CHO HCP
BHK HCP
HEK293 HCP
A549 HCP
MRC5 HCP
PER.C6 HCP

Saccharomyces cerevisiae HCP
Pichia pastoris HCP





SF9 Insect Cell HCP
SF22 Insect Cell HCP





Bovine Serum Proteins
Mycobacterium Proteins
Goat-Milk Proteins




 

Experimental Procedure: HEK293 HCP 2D Western Blot

 

 

 

protein labeling

Protein extraction
Protein assay
Protein labeling

 

HCP 2D gel

2D Gel image:
HEK293 HCP











Gel transfer to Membrane



HCP 2D Membrane


2D Membrane image:
HEK293 HCP





HCP 2D Membrane


2D Membrane image:
Anti-HEK293 HCP Western Blot

 



HCP image overlay


Image Overlay:
Green: HEK293 HCP
Red: Anti-HEK293 HCP Western Blot

 


Spot Matching and Quantitation by DeCyder (100% matching, no ambiguity)


HEK293 HCP Spot Map

HCP Protein

Anti-HEK293 HCP Western Blot Spot Map

HCP Western Blot

Brown: 2051 spots detected by anti-HEK293 HCP antibody
Blue: 149 spots not detected by anti-HEK293 HCP antibody (present in HEK293 HCP image only)
Anti-HEK293 HCP Antibody Coverage: 93.2% of 2200 HEK293 HCP spots


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Example 1: Characterization of Anti-CHO HCP antibody by 2D Western Blot

 

CHO HCP 2D Gel Image

CHO Protein

Anti-CHO HCP 2D Western Blot Image

Anti-CHO HCP Western Blot

Image Overlay: Green: CHO HCP  Red: Western Blot

CHO HCP Anti-CHO HCP image overlay

CHO HCP Spot Map

CHO HCP Protein

Anti-CHO HCP Western Blot Spot Map

Anti-CHO HCP Western Blot

Brown: 2389 spots detected by anti-CHO HCP antibody
Blue: 170 spots not detected by anti-CHO HCP antibody (present in CHO HCP image only)
Anti-CHO HCP Antibody Coverage: 93.4% of 2559 CHO HCP spots

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Example 2: Characterization of Anti-E.coli HCP antibody by 2D Western Blot

 

E.coli HCP 2D Gel Image

E. Coli Protein

Anti-E.coli HCP 2D Western Blot Image

Anti-Ecoli HCP Western Blot

E. Coli HCP 2D Gel Image

E.Coli protein

Anti-E. Coli HCP 2D Western Blot Image

Anti-Ecoli HCP Western Blot

Image Overlay: Green: E. Coli HCP  Red: Western Blot

E.coli HCP Anti-E. Coli HCP image overlay

E.coli HCP Spot Map

E.Coli HCP Protein

Anti-E.coli HCP Western Blot Spot Map

Anti-E.Coli HCP Western Blot

Brown: 2226 spots detected by anti-E.coli HCP antibody
Blue: 31 spots not detected by anti-E.coli HCP antibody (present in E.coli HCP image only)
Anti-E. Coli HCP Antibody Coverage: 99.6% of 2235 E. Coli HCP spots

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Example 3: Characterization of Anti-High Molecular Weight E.coli HCP antibody by 2D Western Blot

 

E. Coli HCP 2D Gel Image

E. Coli protein

Anti-E. Coli HCP 2D Western Blot Image

Anti-E. Coli HCP Western Blot

Image Overlay: Green: E. Coli HCP  Red: Western Blot

E. Coli-HCP Anti-E. Coli HCP image overlay

E.coli HCP Spot Map

E. Coli-HCP protein

Anti-E.coli HCP Western Blot Spot Map

Anti-E. Coli HCP Western Blot

Brown: 933 spots detected by anti-E.coli HCP antibody
Blue: 3 spots not detected by anti-E.coli HCP antibody (present in E.coli HCP image only)
Anti-E.coli High Mw HCP Antibody Coverage: 99.7% of 936 high molecular weight E.coli HCP spots

 

Example 4: Characterization of Anti-Yeast (Saccharomyces cerevisiae) HCP antibody by 2D Western Blot

 

Yeast HCP 2D Gel Image

Yeast

Anti-Yeast HCP 2D Western Blot Image

Anti-Yeast HCP

Yeast HCP 2D Gel Image

Yeast Protein

Anti-Yeast HCP 2D Western Blit Image

Anti-Yeast HCP Western Blot

Image Overlay: Green: Yeast HCP  Red: Western Blot

Yeast HCP Western Blot

Yeast HCP Spot Map

HCP Yeast Protein

Anti-Yeast HCP Western Blot Spot Map

Anti-Yeast HCP Western Blot

Brown: 2400 spots detected by anti-Yeast HCP antibody
Blue: 2 spots not detected by anti-Yeast HCP antibody (present in Yeast HCP image only)
Anti-Yeast HCP Antibody Coverage: 99.9% of 2402 Yeast (Saccharomyces cerevisiae ) HCP spots

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Example 5: Characterization of Anti-SF9 HCP antibody by 2D Western Blot

 

SF9 HCP 2D Membrane Image

Protein 2D Image on Transferred Membrance

SF9 HCP 2D Western Blot Image

SF9-HCP Western Blot

SF9 HCP 2D Gel image

SF9-HCP 2D Gel image

SF9 HCP 2D Western Blot Image

SF9-HCP 2D Western Blot

Image Overlay: Green: SF9 HCP  Red: Western Blot

SF9-HCP Western Blot image overlay

 

SF9 HCP Spot Map


Anti-SF9 HCP Western Blot Spot Map


Brown: 2623 spots detected by anti-SF9 HCP antibody
Blue: 285 spots not detected by anti-SF9 HCP antibody (present in SF9 HCP image only)
Anti-SF9 HCP Antibody Coverage: 90.2% of 2908 SF9 HCP spots

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