+44 191 255 8899 info@totallab.com

The First 2D Gel Analysis Software Exclusively for Host Cell Protein (HCP) Coverage Analysis

Example of 2D Gel Image in Colour Overlay Mode in SpotMap

Developed for:

  • Bioassay Development:

Host cell protein (HCP) and anti-HCP antibody characterization via 2D gels and Western blots.

  • Bioprocess Improvement:

Assess the purification of biologic production through comparison of multiple 2D gels.

  • Analytical Development:

Developing process-specific immunoassays for use in HCP analysis of new Biologics and Biosimilars.

Adopted and trusted globally for analytical development by major Biopharmaceutical, Biotechnology, Chemical companies, leading research organisations and university institutions around the world.

Removal of process-related impurities such as residual host cell proteins (HCPs) to the lowest possible levels within the final product ensures the safety, purity and potency of the final product. Monitoring HCP’s throughout production requires development of specific assays with characterisation of the antigen and antibody.
 
2D gel and Western blot comparison is the current best practice for measuring host cell protein impurities.
 
In addition, to support new marketing applications, establishing sufficient, clear data results and conforming to specifications are critical standards needed to present to regulatory authorities such as the NDA and FDA. Therefore getting some results or using slow, unsuitable tools is not an ideal solution for obtaining reproducible and accurate results.
 
SpotMap is the only 2D gel analysis software created for host cell protein (HCP) coverage analysis that helps to aid your regulatory requirements including 21 CFR Part 11.

Developed for…

  • 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.”

Virginia Liu-Compton

Research Associate 2, ImmunoGen Inc.

Example of 2D Gel Image in Colour Overlay Mode in SpotMap
SpotMap Example Analysis Report using challenging 2D Gel and Western blot images

Reproducible results in less than 1 hour

An example report to illustrate how quickly the software can be used to analyse challenging 2D gels and Western blots and calculate host cell protein 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)Process-related impurities, such as host cell proteins, have set guidance and quality considerations. These include requirements with respect to supported data including 21 CFR part 11.
 

We have taken action to account for these new regulatory requirements following customer feedback on SpotMap and in response we have developed built-in tools to help conformance for your host cell protein analysis.

  • Data Reporting
Export, clear, detailed and complete reports straight out of the software. In addition tools for individual images and tables for reports are available. Allowing full control of your presentation and results.
  •  Customisable Reports
Create clear, complete reports. You can add any additional data, information and images to your reports.
  • Built-in Audit Trail

Generating secure, complete, accurate audit reports is possible with a only a few clicks, helping you focus on your research. These reports enhance transparency, prevent alteration which ensures data integrity. The audit trail generator identifies who, what and when changes have been made in the project.

“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.”

Craig Pattinson

Research Technician, Proteomics, Aberdeen University, The Institute Of Medical Sciences

Need custom-tailored reports?

Host Cell Protein Analysis 21CFR Report

Host Cell Protein Coverage Analysis Example Using SpotMap

A full analysis report and audit trail generated under an hour using challenging gels with SpotMap.

Increased Host Cell Protein Coverage, Accurate, Reproducible Results

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 by enhancing these visual distortions using our unique range of visualisation tools. These tools have been designed to support a reproducible output and have been tested on a variety of representative image sets. Tools include for example: Interactive 3D Editing, Colour Overlay Mode, Secondary Image Overlay.

These methods therefore allow for a more informed and confident decision to be made where necessary and in addition allow an analysis to be reproduced with minimal effort.

“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.”

Virginia Liu-Compton

Research Associate 2, ImmunoGen Inc.

Example of 3D Colour Overlay in SpotMap

Colour Overlay

Example of 3D Secondary Image Overlay in SpotMap

Secondary Image Overlay

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 often restrict your analysis and in addition attempt to predetermine when you should address certain challenges in your analysis.

Comparison of 2D gels and Western blots do not follow such a simple, predictable analysis therefore software must be flexible to adhere to the varied results obtained from this technique.

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”

Rebecca Oettle

Phd Student, University Of Cambridge

Inter- user Variation Poster

Inter-user variation of percentage coverage is within a range of 2% using an optimised analysis process

This report assesses SpotMap across three data sets of varying difficulty, by three users of varying expertise to obtain the host cell protein coverage result. In conclusion, percentage coverage is within a range of 2% using an easy data set and percentage coverage is within a range of 10% using a challenging data set.

Confidently Identify More Host Cell Proteins, Faster

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 consequently make spot detection difficult and in addition decrease the objectivity of the analysis.

SpotMap has highly improved imaging techniques for intuitive automatic spot detection. You can now uncover more low abundance spots and more “hard to detect” spots. Therefore reducing the need for manual detection and again increasing your analysis objectivity thus allowing reproducible results even with challenging data sets.

“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.”

Craig Pattinson

Research Technician, Proteomics,, Aberdeen University, The Institute Of Medical Sciences

Poster: Removal of Positional Variation in SpotMap

Two example methods to removal positional variation in 2D gel and Western blot comparison

The use of additional images to be used as the alignment targets can simplify the alignment process in some cases. However it is not a necessity to complete the analysis. When completing an analysis the approach used should be similar or the same each time a new analysis is used for reproducible results.

Ready to try SpotMap for yourself?

We illustrate the techniques, tools and software best practices so you can overcome your 2D gel and Western blot challenges.

You will learn 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 a 2D gel HCP analysis result and report, including an audit report.

But don’t take our word for it. You will now be ready to test the performance of SpotMap yourself using your own images for free. During which you will have access to our support and analytical team for advice and to answer any technical questions you may have.

Useful Resources

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. Users were in a range of 2% of each other when using easy data sets.

To illustrate, three data sets were used and analysed by three different users with varying range of experience which you can view here. Users analysed SpotMap to determine the relative coverage of the Western blot vs the 2D gel imageWe compared the percentage coverage results between each user.

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 using SpotMap you can overcome these challenges. Features unique to SpotMap enhance these visual distortions allowing a more informed and confident decision when necessary. In addition, tools such as interactive 3D, colour overlay mode and secondary image overlay easily overcome these challenges during your analysis by enhancing your images.

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. Examples illustrated in our evidence page.

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. In addition, you can export individual images and tables from your analysis 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. In fact, alignment is one of the most important features which actually removes positional variation between images, providing easy comparison of spot patterns between images. Spot detection is an intuitive automatic algorithm that identifies the location of spots in the project. This includes those hard to detect spots for example: low abundance spots.

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.