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Pre-clinical and clinical batches

HCP analysis for documentation of pre-clinical and clinical batches

Do you want to identify host cell proteins in your pre-clinical tox batch or GMP phase I/II batches?

We develop sensitive mass spectrometry and gel-based methods; typically used for detection, vizualization and relative quantification of host cell proteins down to low ppm concentrations. We use these methods to identify HCPs in pre-clinical and clinical batches.

Currently, we offer three methods for identifying HCPs:

  • GeLC-MS/MS
  • LC-MS/MS
  • 2D PAGE MALDI MS

The GeLC-MS/MS method is for identification of host cell proteins down to ~50 ppm. We separate the sample by 1D SDS PAGE; each lane is completely excised into 8 gel consecutive pieces and in-gel digested with trypsin. Afterwards, we analyze the peptides by high sensitivity nanoLC-MS/MS and search the data against a customized database containing host cell proteins from your specific cell line and the drug substance sequence. The MS-data is searched against this specifically designed database for HCP identification.

The GeLC-MS/MS analysis provides you with a comprehensive list of observed HCPs, amino acids sequences, theoretical pI/MW, database accession number, etc. The sensitivity of the analysis is in the low-to-sub nanogram range demonstrated on protein standards.

Read more about HCP analysis using GeLC-MS/MS in our poster Combining platform ELISA and orthogonal GeLC-MS/MS for HCP detection, and in the application note Detection of small HCPs by GeLC-MS/MS.

Download application note in low resolution
(422 KB)

Download application note in high resolution
(4.6 MB)

The LC-MS/MS method with SWATH data acquisition is for identification and relative quantification of HCPs down to low ppm concentrations. We digest the drug substance in-solution and then analyse the complex peptide mixture by LC-MS/MS. Our scientists perform this analysis on a Sciex TrippleTOF 6600 instrument using data-independent SWATH data acquisition and ion library searching. The analysis is under development, promissing to deliver low-to-sub-ppm detection limits and highly reproducible quantification of each HCP.

The 2D PAGE with MALDI MS method is for visualization and identification of proteins separated by 2D PAGE according to pI and Mw. We stain the proteins by coomassie blue or fluorescense, then cut out the visible spots for in-gel trypsin digestion and MALDI MS/MS peptide analysis of each spot. The HCP protein identification is obtained by a MASCOT search in customized HCP protein database. Typically it identifies >95% of visible protein spots. The sensitivity of the analysis is demonstrated on known protein standards.

We also develop host cell protein analysis for up- and downstream bioprocess optimization

 

Services/Products
Protein analysis by Alphalyse

Identification of host cell proteins by GeLC MS/MS

  • Identification of host cell proteins
  • 1D PAGE nanoLC-MS/MS
  • Sensitivity ~50 ppm
  • HCP list with pI, Mw, aa sequence, acc number

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote
Protein analysis by Alphalyse

Identification of host cell proteins by in-solution digest & SWATH LC MS/MS

  • Identification of host cell proteins
  • SWATH LC-MS/MS analysis on Sciex TrippleTOF 6600 mass spectrometer
  • Database search in custom HCP Ion library
  • HCP list with pI, Mw, aa sequence, accesion number

$12,000.00

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote
Protein analysis by Alphalyse

Identification of host cell proteins by 2D PAGE & MALDI MS/MS

  • Identification of host cell proteins
  • 2D PAGE MALDI-MS/MS
  • Sensitivity ~all visible spots
  • HCP list with pI, Mw, aa sequence, acc number

$12,000.00

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote

Technical details

Sample requirements

Sample shipment

shipping option
Ship your protein samples by a courier company of your own choice

Place your samples into micro tubes and pack in a suitable transport box.

  • Contact a courier company of your choice (FedEx, UPS, DHL, TNT, World Courier etc.). Use the following sentence to describe the content: “Protein material – Non-toxic, non-hazardous, for research purpose only“.
  • Use cool packs or cooling units if your samples should be stored below 5°C/40°F
  • Ship the samples to the office closest to you (Canadian customers, please ship to Europe):
Customers in the USA
Alphalyse, Inc.
200 Page Mill Road, Suite 100
Palo Alto, CA, 94306
USA
Att: Jette Williams
Tel: (650) 543-3193
Customers outside of USA
Alphalyse A/S
Unsbjergvej 4 D
DK-5220, Odense
DENMARK
Att: Anette Rasmussen
Tel: +45 6310 6500
shipping option
Order a prepaid FedEx Air Waybill < 1 lb./ 500g

A prepaid FedEx Air Waybill can be used for fast and easy shipment of samples in packages up to 1 lb./ 500g.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack in suitable transport box.
  • Print the PDF document and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to send your samples stored at room temperature, this option is usually the right choice. If you fill your box with cooling packs, the weight will most often exceed 1 lb./500g.

shipping option
Order a prepaid FedEx Air Waybill < 10 lb./5kg (e.g. for packages with cooling packs)

This prepaid FedEx Air Waybill can be used for shipment of samples in packages up to 10 lb./ 5kg, with or without cooling packs.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack them in a suitable transport box. Fill with cooling units/cold packs.
  • Print the PDF and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to fill your transportation box with cooling packs, select this shipping option.

shipping option
Get a quote for shipment by World Courier

The World Courier service can be used for shipment of samples of all types. If you select this option, we will contact you to provide a quotation based on location and shipping conditions. You will receive a prepaid Air Waybill which will be invoiced together with your analysis results. This is usually a good option if FedEx does not service your location.
With the prepaid World Courier service you get:

  • GxP compliant processes to ensure maximal security and traceability
  • Door-to-door solution on a dedicated vehicle
  • Possibility of providing qualified packaging, temperature monitoring and dry ice
  • Shortest possible transit-time through “next flight out”-procedure
  • Customs clearance and help with paperwork

Read more
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