The First 2D Gel Analysis Software Exclusively for Host Cell Protein (HCP) Coverage Analysis
Characterize host cell proteins (HCP) and anti-HCP antibodies via 2D gels and Western blots.
Assess the purification of biologic production through comparison of multiple 2D gels.
Developing process-specific immunoassays for use in HCP analysis of new Biologics and Biosimilars.
Adopted and trusted by some of the major Biopharmaceutical, Biotechnology, Chemical companies, leading research organisations and university institutions around the world.
Process-related impurities such as residual host cell proteins (HCPs) must be removed to the lowest possible levels within the final product to ensure the safety, purity and potency of the product. Monitoring HCP’s throughout production requires development of specific assays with characterisation of the antigen and antibody.
2D gel and 2D Western blot comparison is the current best practice for measuring host cell protein impurities.
Establishing sufficient, clear data results and conformance specifications to support new marketing applications are a critical standard that will be presented to regulatory authorities such as the NDA and FDA. Getting some results, or using slow, unsuitable tools from your current analysis solution is not an ideal solution for obtaining reproducible and accurate results.
SpotMap is the only 2D gel analysis software that has been created exclusively for host cell protein (HCP) coverage analysis that helps to aid your regulatory requirements including 21 CFR Part 11.
- 2D gel and 2D Western blot,
- 2D Differential in Blot Electrophoresis (2D-DIBE),
- 2D Fluorescence Difference Gel Electrophoresis (2D-DIGE), using any stain including CyDyes
“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. […] With this software, my team was able to make a more informed choice on the best candidate to use for our purposes.”
Reproducible results in less than 1 hour
Using challenging images with SpotMap discover how quickly the software can be used to analyse 2D gels and Western blots and calculate percentage coverage.
View a small sample of reports, posters, application notes and more demonstrating the use of SpotMap.
Secure Reliable Results. Helps To Support 21 CFR Part 11
New regulatory actions have set forth minimum requirements to assure that drugs meet the standards of the Federal Food, Drug, and Cosmetic Act (FD&C Act) this includes requirements with respect to data including 21 CFR part 11.
SpotMap is the only analysis software that has taken into account these new regulatory requirements and has built-in tools to help conformance.
- Data Reporting
You can use our automatic report generator to export a clear, detailed and complete report straight out of the software. Individual images and tables can be exported separately for reports and presentations, giving you full control of your presentation report and results.
- Customisable Reports
If desired you can input any additional data, information and images to your report that may help make them more readily available to external regulators and auditors.
- Built-in Audit Trail
Secure, accurate generation of a complete audit trail. These static electronic records identify who, what and when changes have been made in the project. This enhances transparency, prevents alteration which ensures data integrity. These record-keeping procedures are now a minimum requirement to assure that drugs meet the regulatory standards set by the FDA.
“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). The best part is I can depend on the results.”
HCP Coverage Analysis Example
Using challenging images, a full analysis was performed and a report was generated including an audit trail to support 21 CFR Part 11 compliance in under an hour using SpotMap.
Greater Coverage Accuracy, Reduced Subjectivity
Complex sample preparation and running procedures cause distortions such as faint spots, overlapping spots, saturated spots and streaks making analysis difficult.
You can overcome these challenges during your analysis easily by enhancing these visual distortions using our range of visualisation tools, allowing a more informed and confident decision where necessary. Tools include interactive 3D, colour overlay mode and secondary image overlay.
Secondary Image Overlay
“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.”
Reduce Analysis Time, One Dedicated User Interface
Positional variation, gel distortion and image quality are all demanding challenges when comparing 2D gels and Western blots. Software
workflows restrict your 2D analysis and attempt to predetermine when you should address those challenges.
SpotMap puts you back in control of your analysis with a single flexible user interface optimised for 2D gel and Western blot comparison – including single stain, 2D-DIGE, 2D-DIBE and AAE experiments. It combines all the proven, objective features into one customisable workflow to satisfy all variations of 2D gels and Western blots.
“I have found the software very easy to use and has been exactly what I needed to compare western blots and pick the spots from my gel”
Inter-user variation of percentage coverage is within a range of 2% using an optimised analysis process
SpotMap was assessed across three data sets of varying difficulty, by three users of varying expertise to obtain the HCP coverage result.
Improved Spot Detection, Minimal User Intervention.
Low abundance spots, distorted spots, overlapping spots, spots suffering from noise, saturated spots, streaking and the presence of non-spot features are common issues faced in 2D analysis and make spot detection difficult and decrease the objectivity of the analysis.
SpotMap has highly improved imaging techniques for intuitive automatic spot detection. Uncover more low abundance spots, reduce the need for manual detection and increase your analysis objectivity.
“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.”
Removal of positional variation comparing two different alignment targets
In this analysis the use of two different alignment targets were compared. In one analysis the Western blot was aligned to the gel. In the other analysis, both the 2D gel and Western blot were aligned to the stained membrane of the Western blot. 1455 spots were detected using automatic detection on the directly aligned image compared to 1558 spots following indirect alignment.
Ready to try SpotMap for yourself?
We can jump online with you for an hour and demonstrate live how SpotMap can easily overcome your 2D gel and Western blot challenges. The demonstration consists of the following insights:
- How you can achieve reproducible results
- How you can overcome analysing typical challenging 2D gel and Western blot images.
- Which analysis tools you should be using to employ best practices.
- How you can quickly produce and export a report and audit trail.
After which you can be up and running with a trial of the software to analyse your own images for free for up to two weeks.
The software is going to be used in a cGMP lab. Is SpotMap suitable for the regulatory requirement including 21 CFR?
SpotMap now has new tools which help compliance with 21 CFR part 11 and is now the only software developed for immunoassay development, analytical development and bioprocess improvement that aims to support new regulatory requirements.
What images can I use with SpotMap?
SpotMap is compatible with most imaging scanners (e.g Amersham Typhoon system, GE Typhoon) and supports most image formats produced from laser scanners including jpg, png, gel, mel as well as 8-bit and 16-bit TIFF files. For further technical details you can contact us directly here.
How much variability would you see between users?
Results obtained from our reports showed users were within a range of 10% of each other, when using the most challenging images. Using easy data sets a range of within 2% were obtained.
In this report which you can view here, three data sets were used and analysed by three different users with varying range of experience. Each data set was analysed using SpotMap to determine the relative coverage of the Western blot vs the 2D gel image. For each data set the percentage coverage results were compared between users.
Typically with 2D gels and 2D Western blots the biggest source of variation is the quality of the image used. The use of high quality images increase the objectivity and reliability in your results. Effects such as colour depth, saturation, streaking and presence of non-spot features reduce this objectivity. However SpotMap was designed to overcome these challenges. Features unique to SpotMap enhance these visual distortions allowing a more informed and confident decision when necessary. Tools such as interactive 3D, colour overlay mode and secondary image overlay easily overcome these challenges during your analysis.
How long does an analysis usually take?
Multiple studies show that users can analyse their 2D gels and Western blots and produce objective results within one hour, even using challenging images. Go to SpotMap evidence page to view a small portion of these studies.
I want to easily be able to present my results and processes to external regulators, can I do this in SpotMap?
The built-in report generator button exports your whole analysis in a clear, detailed pdf report and is the fastest way for third party regulators to view your results. You can export individual images and tables if necessary,
The audit-trail exporter ensures accurate generation of a complete audit trail . These static electronic records identify who, what and when changes have been made in the project. This enhances transparency, prevents alteration which ensures the records exported are the original records and results.
How can I ensure my analysis is reproducible?
Automatic tools in SpotMap increase your reproducibility such as automatic spot detection and automatic alignment. Alignment is a key feature which removes positional variation between images, providing easy comparison of spot patterns between images. Spot detection is an automatic algorithm that identifies the location of all spots in the project.
16 bit images are recommended for analysis as these increase the intensity scale from 0-65535. You can find more advice and best practices required to generate digital images of a quality suitable for automated image analysis here. However SpotMap can analyse most image formats including 8 bit TIFF and lossy image formats.