HCPs in Biosimilar vs Originator

  • Identify and quantify all HCPs in drug substance
  • Compare HCP levels in originator to multiple biosimilars
  • Get regulatory documentation for FDA/EMA approval

Do you want to compare process-related impurities in biosimilar and reference products?

For approval of originator products, a comprehensive documentation of host cell proteins (HCPs) must be carried out. Even though biosimilars contain the same active product ingredient (API), the HCP content might not be the same.

Thus, the API in biosimilars might have different activity or immunogenic ability, and should be subject to ...

Do you want to compare process-related impurities in biosimilar and reference products?

For approval of originator products, a comprehensive documentation of host cell proteins (HCPs) must be carried out. Even though biosimilars contain the same active product ingredient (API), the HCP content might not be the same.

Thus, the API in biosimilars might have different activity or immunogenic ability, and should be subject to just as extensive an HCP analysis as their originators.

We can provide you with a side-by-side comparison of process-related impurities compared to originator. For a sensitive and reproducible analysis of HCPs in originator and biosimilar we perform the analysis by mass spectrometry.

Note, this LC-MS analysis does not require access to ELISA reagents.

This service includes

  • A generic sample preparation for purified batches and process samples in different matrices
  • LC-MS peptide analysis on a robust microflow HPLC and Sciex TrippleTOF 6600 mass spectrometer
  • SWATH LC-MS for reproducible HCP identification and quantification
  • A report with lists of HCPs and their physiochemical properties (MW, pI, AA sequence, database accession number) for both biosimilar and originator
MoreLess details

The process:

  • 1 Contact us to discuss your project and receive a project proposal
  • 2 Analysis phase lead by Alphalyse appointed principal investigator
  • 3 Report w. overview of HCP content, compared between originator and biosimilars
  • 4 Follow-up by phone or email

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We offer customized solutions, contact us to discuss your project.

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Testimonials

  • "The most sensitive, yet robust, Host Cell Protein analysis we’ve seen"

    Analyses of key protein features, stability and impurity levels

    See more
  • "Alphalyse´s HCP analysis saved us the development of an ELISA assay that may not have worked anyway."

    Innovative development of an orthogonal method for Host Cell Protein analysis

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Knowledge center

More information

We would like to help you as much as possible with your project and therefore provide several kinds of customer support:

Technical details

What is SWATH?

The SWATH mass spec technology is a data independent acquisition strategy. So this means that we collect all MS/MS fragmentation data for every peptide in your sample. This is only possible due to the high speed of modern mass spectrometers.

The Sciex 6600 TripleTof instrument makes it possible to divide the whole mass range into sequential windows of size from 5-50 daltons. All the while we are still able to perform MS/MS fragmentation covering the full mass range (m/z 350-1700) within a few seconds. We set up the individual windows based on analysis of your samples, and then typically get 50-100 windows.

In this way we obtain MS/MS data for all peptides without having the instrument select certain peptides for MS/MS fragmentation.

Quality control standards

Standard curves for quality control of biosimilar products

 

The MS/MS data set links to the retention time and can contain MS/MS data from multiple peptides. A spectra library, linking to peptide sequences in a library, extracts the MS/MS data.

The spectra library can be prepared in many ways. The most simple way is to run a few samples in data dependent acquisition mode to collect good MS/MS data. Or a more advanced way is basing it on off-line fragmentation followed by MS/MS analysis of each fraction. The library can be extended with new MS/MS data at any given time, and used for a research of old SWATH data.

Sample preparation

Sample requirements for SWATH LC-MS/MS

HCP analysis using SWATH technology requires sample material in sufficient amounts to obtain good data. Also, it is important that samples are prepared in a clean laboratory to avoid contamination with human keratin.

Samples can be submitted in liquid or lyophilized form.

Samples for Library generation and SWATH analysis

  1. The analysis requires typically a minimum of 500-1000 μg of protein material for each sample.
  2. The samples should not contain large amounts of detergents.
  3. The concentration should be >0.5 µg/µl.
  4. Use a Eppendorf Safe Lock tube or similar tubes from your lab.
  5. Freeze or refrigerate to +4 oC for cold shipment of the liquid sample.

Meet the experts

For this type of analysis our experts include:

Do you need help?
Rikke Raaen Lund

PhD in Biomedicine

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Do you need help?
Janne Skaarup Crawford

BSc in Cell Biology and Biochemistry

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Do you need help?
Inga Boll

MSc in Biochemistry & Molecular Biology

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