SpotMap - Host Cell Protein (HCP) Coverage % Analysis Software

A complete solution for analysing HCP coverage using 2D gel electrophoresis and Western blot images

Our SpotMap software first launched to market in 2015 and is the industry-leading software for host cell protein (HCP) antibody coverage analysis in the pharmaceutical industry.

SpotMap is a complete solution for analysing HCP coverage with 2D gel electrophoresis and Western blot images.

  • Universal. Supports all industry-standard image formats, including TIFF, GEL, MEL and IMG files. It also supports single stain, DIGE and DIBE techniques.
  • Speed. Get a coverage % in less than 10 minutes.
  • Objectivity. Unmatched automation reduces the need for manual intervention.
  • Reproducible. Evidence of consistent coverage results between different users and labs, drastically reduce subjectivity in your analysis.
  • Quantitation. Enables reliable, accurate quantitation of gel and blot images.
  • 21 CFR/GMP compliant Meet FDA 21 CFR part 11/EU Annex 11/GAMP5 GMP compliance for compliant labs, from captured image through to final analysis and report production.

SpotMap is unique and easy-to-use software specifically designed to automate the process of analysing HCP (host cell protein) coverage for biologic product characterisation, process development/optimisation and HCP antibody assay & impurity profiling.

The software has been developed in line with and in response to pharmaceutical industry feedback, it is easy to master, with accurate results obtained quickly and reproducibly.

Our SpotMap software is trusted by the world’s leading pharmaceutical/CDMO companies

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"SpotMap played an integral role in our comparison of proteomes of different cell lines during the process of choosing a proper antigen for antibody development. The ability to overlay gels and assign numbers to each protein spot was especially useful in determining similarities and differences between gels without duplicating identified spots. One feature that I found to be especially useful was the 3D view which allowed for a better visual of spot intensity when determining spots. The table of identified spot for each image also made it easy to identify which spots are unique or common to the different images. With this software, my team was able to make a more informed choice on the best candidate to use for our purposes."

Virginia Liu-Compton
- Senior Scientist II, ImmunoGen Inc, Waltham, Massachusetts, USA

Why do I need to know my HCP coverage?

SpotMap is designed to help users analyse HCP coverage – the percentage of immunodetection that an antibody reagent covers for the total population of HCPs. It does this by comparing a 2D gel or blot image of proteins detected with an anti-HCP antibody (your secondary image) against an image stained to reveal all possible HCP antigen proteins (your primary image).

HCP coverage analysis is used for:

  • Development of analytical methods (choice and validation of immunoassay reagents)
  • Selection and validation of the most appropriate generic HCP ELISA kit
  • Bridging studies to support assay replacement (e.g. change from commercial assay to platform assay or process-specific assay)
  • Generic assay re-validation for every new batch of reagent
  • Process- and platform-specific assay revalidation after changes that could affect HCP composition
  • Optimization of bioprocesses

2D electrophoresis and Western blotting remains the gold standard for orthogonal anti-HCP characterisation as outlined in the U.S. Pharmacopeia National Formulary:

We have had real difficulty matching reactions on blots to the corresponding areas on the gels. With our previous technology it genuinely wasn’t possible so adding the SpotMap software to our facility has helped greatly. […] Once we had the images put through SpotMap software it corresponded to areas that personally I would have never matched, which ended up being the correct areas.

Craig Pattinson
- Research Technician, Proteomics Aberdeen University

Optimise your host cell protein analysis workflow

We use intuitive, automated algorithms and an easy-to-use interface to enable users to obtain an accurate coverage percentage in around 10 minutes.

This enables users to achieve extremely reproducible antibody-to-host cell protein coverage, even between different users and labs. It provides reliable, accurate quantitation of gel and blot images so that you can achieve objective results.

Users are also able to verify host cell protein antibody reactivity, giving you a more complete understanding of the antigen quality.

If you’re looking for software to support HCP coverage analysis, make bioprocess improvements or achieve analytical development then request a free demo of SpotMap. You’ll receive guidance on how to achieve reproducible results, tips for aligning challenging gels and Western blots and knowledge on the software tools to use on more challenging images, such as faint spots.


Report with confidence

  • Automatically generate a comprehensive report of your entire analysis, including original images analysed, summarising all essential quality control checks, analysis parameters, results, images, tables and coverage charts in a single document. A detailed spot measurements table can also beappended to your report
  • Easily visualize and export critical analysis parameters such as position of alignment vectors and spots that were excluded or edited
  • Save, print or copy to clipboard all project data, tables and images (2D and 3D views, graphical reports, Display area, Application window)
  • Carry out pI/MW calibration to help interpretation of mass spectroscopy-based identification data

FDA 21 CFR part 11 regulations stipulate the need to track, record and authenticate your experiments. With our GxP Module, you’re able to monitor system access, record user activity and define project sign-off procedures to ensure compliance with FDA regulations. You can read more about it here.

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Case Studies

Case Study: Western Blot Assessment of Polyclonal Anti-Host Cell Protein Antibody Production, Covance BioCMC (Now LabCorp)

Case Study: University of Aberdeen, Proteomics Core facility

“The proteomics core facility provide protein analytical services to academics and companies, mostly in the local area but also worldwide. We offer a wide range of services to try and accommodate any needs, these can vary from simple protein identifications to large scale gel/mass spec experiments. Currently we are mainly offering services in: Gel electrophoresis (1D, 2D, DIGE), In-depth proteome analysis (Q-Exactive LC-MS), molecular histology (MALDI- imaging), protein identification/quantification, mapping of protein modifications and rapid microbial identification (MALDI Biotyper).”

“We have had real diffculty matching reactions on blots to the corresponding areas on the gels. With our previous technology it genuinely wasn’t possible so adding the SpotMap software to our facility has helped greatly. Because a big reaction on blots doesn’t necessarily mean a big amount of protein on the gel it’s sometimes hard to match patterns by eye. In the past we have had to rely on matching patterns by eye and end up cutting out the wrong area of the gel. Once we had the images put through the SpotMap software it corresponded to areas that personally I would have never matched (which ended up being the correct areas).”

Craig Pattinson, Research Technician, Proteomics, Aberdeen University

Case Study: Rockland Immunochemicals

Poster, originally presented at the WCBP 2016 conference: “2D-SDS PAGE Analysis Methods For Determining HCP Coverage Using Process Derived anti-HCP Polyclonal Antibodies”

“Both spot analysis methods generated a final coverage that was between 44 – 59%. However there was much more variation in coverage between the 1.0 second exposure and 5.5 second exposure (11%) using the PD Quest Spot analysis. Spot analysis using SpotMap resulted in coverage that was consistent within 1% between all three blot exposure times. Workflow between the two analysis systems varied by about 2 hours – analysis took approximately 1 hour using the SpotMap software and approximately 3.5 hours using the PD Quest software. PD Quest spot analysis software offers numerous options and parameter selection criteria whereas SpotMap software has limited parameter selection but requiring fewer steps in the workflow to determine antibody coverage.”

Case Study: Minimising Inter-User Variation in Coverage of 2D Gels

This report demonstrates the inter-user variation of SpotMap used by three individuals of varying experience on three data sets categorised as hard,
medium and easy. Results obtained were within a range of 10% of each user, even when the most challenging images were used. Key areas of the analysis
where variation may be introduced have been identified.

“Typically with 2D gels and Western blots the biggest source of variation is the image quality. The automatic image QC flagged two of the data analysis results as being potentially imprecise due to the use of 8-bit images. Smearing/steaking, saturation and presence of non-spot features also affects analysis precision.

There are areas of the analysis that require particular consideration to improve objectivity and precision. Within the software, image editing, alignment vectors added, spot detection parameters used and individual decisions on the presence of a spot will all impact the results.

However, SpotMap gives reproducible and reliable results within a range of 10%, even using challenging images. The use of high quality images which enable a consistent analysis approach between users will reduce the variation, as seen in the easy data set where results are within a range of 2%. Production of an SOP that addresses all aspects of the analysis, including optimised gel running and image capture would increase the objectivity of the results, reducing the subjective decisions required during analysis.”

Case Study: Rockland Immunochemicals

Poster, originally presented at the WCBP 2018 conference: “Image analysis used to minimise inter-user and inter-lab variation of results
from measuring HCP-antibody coverage by comparing features between 2D gel and 2D Western blots.”


Our SpotMap software has been in active development since 2011 and in that time has been cited numerous times in proteomics and anti-HCP antibody generation publications within the field.


You can find a comprehensive list of papers published using SpotMap here.