Protein analysis by UV HPLC

  • Characterization of unknown peaks and shoulders
  • Stability study
  • Custom method setup

Determine the liquid chromatographic pattern of  proteins in drug sample

UV HPLC analysis is an important part of protein characterization. The chromatography may leave you with questions about the identity of specific peaks and shoulders in your chromatogram.

However, a molecular weight analysis of the peaks by mass spectrometry will often identify the molecule.

We can help with:

  • ...

Determine the liquid chromatographic pattern of  proteins in drug sample

UV HPLC analysis is an important part of protein characterization. The chromatography may leave you with questions about the identity of specific peaks and shoulders in your chromatogram.

However, a molecular weight analysis of the peaks by mass spectrometry will often identify the molecule.

We can help with:

  • Standard or customized HPLC analysis of your sample
  • Collection of fractions for further analysis
  • Combination with ESI-MS analysis for mass determination of each peak, N-terminal Edman Sequencing and peptide mapping analysis

Do you need to set up a custom HPLC analysis method?

You can also ask us to customize a method that meets your specific needs. For instance an in-house method, which you want us to set up in our laboratory. We can combine it with high resolution Mw determination by mass spectrometry.

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The process:

  • 1 Contact us to discuss your project and receive a project proposal
  • 2 Analysis phase, where a project leader is appointed to manage the project
  • 3 You receive the report within a few weeks, depending on the analysis
  • 4 Follow-up by phone or email

Start the collaboration

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

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Testimonials

  • "They quickly presented an optimized method that provided excellent data and clearly separated product related impurities."

    Optimized HPLC analysis of peptides for clinical trials and stable GMP production

    See more
  • "We are very pleased Alphalyses high-quality tests. We can ship them hundreds of samples at once and always receive the analytic results shortly after."

    High-throughput characterization analysis of antibodies for fast quality assessment

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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

Introduction to UV HPLC analysis

The analysis is conducted on our Agilent 1290 HPLC systems equipped with fraction collector, UV and fluorescence detector.

Standard analysis

We have set of standard analysis including column and gradient settings.

Custom designed analysis

The custom designed analysis is based on HPLC settings from clients including e.g. column, temperature and gradient settings. The analysis can be coupled to MS instrument to obtain accurate mass information of all peaks.

 

HPLC chromatogram with protein mixture

UV-HPLC Chromatogram of a protein mixture

Sample preparation

Sample requirements for UV HPLC analysis

Protein samples can be submitted in liquid form or as lyophilized material.

Minimum amount of material ~ 20-50 micrograms.

Avoid detergents and keep buffer concentration at a minimum. HPLC analysis may work on samples containing small amounts of salts or urea. However, the best results are obtained with low buffer strength in volatile solvents without detergents.

  • Put the sample into microcentrifuge tube
  • Use Eppendorf Safe Lock tubes, or similar tubes from your lab
  • Lyophilize or speed vac the sample to a solid to ensure protein stability during shipment
  • Alternatively, freeze or refrigerate to +4°C for cold shipment of the liquid sample
Related services

Meet the experts

For this type of analysis our experts include:

Do you need help?
Martin Villadsen

MSc in Engineering within Medical Biotechnology

Read more
Do you need help?
Fen Yang Reske-Nielsen

PhD in Protein Chemistry

Read more
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