HCP Analysis: Antibody Coverage

HCP Analysis: Detect And Quantitate 3 HCP Samples In 1 Test
HCP Analysis: Product Purity & HCP Contaminant

Applied Biomics offers the most comprehensive HCP Antibody Coverage analysis by using high resolution large format 2D fluorescent Western Blot. Our report provides HCP, Western blot and HCP/Western overlay images, allowing the direct visualization of antibody coverage that no other platform could achieve. In addition, the HCP antibody coverage is calculated with high consistency and accuracy (see details below). This platform has been successfully applied in the following areas:

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

Our HCP Antibody Coverage platform offers significant ADVANTAGES over ELISA and standard 2D Western blot:

1) High consistency: Fluorescent 2D Western detects both protein and WB images from the same gel and same membrane, allowing the perfect alignments between protein and WB spots, thus eliminating any gel-to-gel or gel-to-membrane variations as seen in the traditional 2D WBs.
2) Cost-effective: Since only one gel is needed, it saves the cost of running duplicate gels.
3) High accuracy: Fluorescent CyDye labeling allows accurate quantitation of protein and WB spots.
4) High sensitivity: Fluorescent CyDye labeling has a higher sensitivity and wider dynamic range over the silver staining.

HCP 2D Western Blot Experimental Procedure
Example-1: CHO Cell HCP Antibody Coverage
Example-2: E.coli HCP Antibody Coverage
Example-3: E.coli High Mw HCP Antibody Coverage
Example-4: Yeast (Saccharomyces cerevisiae) HCP Antibody Coverage
Example-5: SF9 HCP Antibody Coverage

Click to view a complete list of Applied Biomics HCP assays

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 extraction Protein assay Protein labeling
		2D Gel image: HEK293 HCP
		Gel transfer to Membrane
		2D Membrane image: HEK293 HCP
		2D Membrane image: Anti-HEK293 HCP Western Blot
		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	Anti-HEK293 HCP Western Blot Spot Map
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

Example 1: CHO Cell HCP Antibody Coverage by 2D Western Blot

CHO HCP 2D Gel Image	Anti-CHO HCP 2D Western Blot Image
Image Overlay: Green: CHO HCP Red: Western Blot
CHO HCP Spot Map	Anti-CHO HCP Western Blot Spot Map
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

Example 2: E.coli HCP Antibody Coverage by 2D Western Blot

E.coli HCP 2D Gel Image	Anti-E.coli HCP 2D Western Blot Image
E. Coli HCP 2D Gel Image	Anti-E. Coli HCP 2D Western Blot Image
Image Overlay: Green: E. Coli HCP Red: Western Blot
E.coli HCP Spot Map	Anti-E.coli HCP Western Blot Spot Map
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

Example 3: E.coli High Mw HCP Antibody Coverage by 2D Western Blot

E. Coli HCP 2D Gel Image	Anti-E. Coli HCP 2D Western Blot Image
Image Overlay: Green: E. Coli HCP Red: Western Blot
E.coli HCP Spot Map	Anti-E.coli HCP Western Blot Spot Map
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: Yeast (Saccharomyces cerevisiae) HCP Antibody Coverage by 2D Western Blot

Yeast HCP 2D Gel Image	Anti-Yeast HCP 2D Western Blot Image
Yeast HCP 2D Gel Image	Anti-Yeast HCP 2D Western Blit Image
Image Overlay: Green: Yeast HCP Red: Western Blot
Yeast HCP Spot Map	Anti-Yeast HCP Western Blot Spot Map
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

Example 5: SF9 HCP Antibody Coverage by 2D Western Blot

SF9 HCP 2D Membrane Image	SF9 HCP 2D Western Blot Image
SF9 HCP 2D Gel image	SF9 HCP 2D Western Blot Image
Image Overlay: Green: SF9 HCP Red: Western Blot
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 If you are interested, please follow these steps to obtain more information and order this service.