face

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

ID 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
Order analysis now
Newsletter Subscribe to the Alphalyse Newsletter to get 3 tips
about protein analysis methods and applications
Private, secure, spam-free
© Copyright Alphalyse , Inc. 2002 - 2017. All rights reserved.