Fast and reproducible 2D gel and Western blot analysis software for anti-HCP antibody characterisation.


Why SpotMap?

SpotMap is a unique and easy-to-use software specifically designed for analysis of HCP coverage for biologic product characterisation, process development/optimisation and HCP antibody assay & impurity profiling.

Analysis of HCP coverage is essential for ensuring that biologic products comply to regulations:

  • “Biologic product (…) is safe, pure, and potent” 42 USC 262.
  • “Process related impurities [including HCP] (…) should be minimised” ICH Q6B
  • “Critical Quality Attributes (CQA) [including HCP] (…) should be within an appropriate limit, range or distribution to ensure the desired product quality” ICH Q8.

Current best practice for analysis of HCP coverage is through comparison of 2D gels vs 2D Western blots, using process specific anti-HCP antigens. SpotMap provides the analytical detail required to help you meet regulatory requirements, now and for the foreseeable future.

What is SpotMap?

SpotMap overcomes the challenges you face in analysing 2D gels and Western blots.

SpotMap measures HCP coverage by calculating the relative percentage of protein present in the Western blot compared to the 2D total protein gel.

Images are automatically quality checked and visual issues are clearly identified when uploaded to the software, giving you confidence in your results.

Alignment allows corresponding spots  and blot features to be identified in the same coordinate space.

Spot detection is automated, reducing the subjectivity of results.

Reproducible and reliable results can be easily obtained

The software is easy to use with a simple and quick workflow, analysis of HCP coverage can be completed in less than one day!

“We’ve been looking at different solutions for three months now and from what we just saw [demonstrated] this is already the best”

Major pharmaceutical company following a first demo of SpotMap

“The anti-HCP antiserum needs to be qualified for its ability to detect potential HCP impurities. This assessment should include 2D SDS-PAGE gels of the range of HCPs detected by a sensitive protein stain, such as silver stain, compared to the range detected by western blot analysis using the antiserum employed in the assay.”

OBP Recommended Practice (OBP-RP-004)

 Evaluate SpotMap using your own images

Fill out the form below and in 3 simple steps we will show you how SpotMap can be used to analyse your own images.

  1. Get a personal upload link – we’ll send you a personal log-in and password to our image upload service to the details provided.
  2. Upload your images – follow the simple steps on the upload page to submit your images. We’ll analyse them and alert you when your results are available.
  3. View your results – Download and share your PDF report of your image analysis.

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Examples and evidence of SpotMap in use

Eg Report
SpotMap analysis of challenging 2D gel vs Western blot images. This report demonstrates the simple workflow of SpotMap to determine the relative coverage of blot vs gel based on spot number. In this analysis coverage was 18%. Read more…
Interuser report
Inter-user variation between three users was challenged: results were within a range of 10%. Using low quality images for analysis is a key source of variation as it increases the subjectivity of the analysis. Read more…

Why choose SpotMap?

SpotMap is unique, it is the only software which has been developed specifically for the analysis of 2D gel vs Western blot and for anti-HCP antibody characterisation.

Other software packages exist which could be manipulated to complete this analysis, however these have been developed for the analysis of differential expression not HCP characterisation.

SpotMap is also unique in comparison to using a contract research organisation (CRO). No CRO has SpotMap software, many use analysis packages developed for differential expression or may even analyse the images by eye!

“Workflow between the two analysis systems varied by about 2 hours – analysis took approximately 1 hour using the Spot Map software and approximately 3.5 hours using the PD Quest software.”

Rockland Immunochemicals – 2D-SDS PAGE Analysis Methods For Determining HCP Coverage Using Process Derived anti-HCP Polyclonal Antibodies.

Simple 5 step workflow


The simple workflow of SpotMap allows objective, reproducible analysis to be completed quickly.

1. Images: Each image has a quality check that looks at a range of features. This gives an instant visual check and confidence in the data being analysed.
Image QC
2. Alignment: Alignment makes it possible to accurately compare images by removing the positional variation introduced during the experimental process. 
3. Spots: A master spot map is created which identifies the location of all spots present on all images in the analysis.
Detected spots
3. Match Spots: The master spot map is overlaid on each image in turn and spots are categorised as present or absent on each image.  
5. Results: Results of the whole analysis, 1 vs 1 comparisons and selected areas of the spot map are presented. Drag and drop results and data directly into reports and presentations from the Data Access window. 
Results screen

Technical specification

SpotMap gives you the features you need for 2D gel vs Western blot analysis.

Can I calculate spot coverage?

Yes. A spot map is created of the fully resolved host cell protein, determining the presence of spots expressed on the blot is then used to calculate the relative coverage of Western blot vs the 2D gel.

Can I check my image quality?

Yes. When adding images to the experiment they are automatically checked for quality and any issues/notes are highlighted for you.

Can I edit my images?

Yes. If your images require minor changes (flipping etc.) then you can edit them within the software without changing the original images.

Can I account for 2D gel/Western blot distortions?

Yes. Alignment makes it possible to accurately compare images by removing the positional variation introduced during the electrophoresis and blotting process.

Can I export my results?

Yes. All the calculations, spot numbers, images and data can be easily exported from the program.

Evaluate SpotMap with your own images and get a personalised analysis report

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