HCP Analysis by Mass Spectrometry

Our LC-MS/MS based Host Cell Protein (HCP) analysis uses the Orbitrap Exploris with BioPharma Finder for identification and absolute quantitation of both drug substance (DS) and HCP contaminants at 1-10 ppm levels in the product. We have optimized our work flow starting from sample preparation, mass spectrometry scan settings to data analysis, specifically:

  • Sample Preparation:
    • Optimized HCP sample prep protocols for different HCP sample types
    • Advanced HCP Sample prep: Protein fractionation and/or DS removal to enhance the detection of low abundance HCPs
    • See more customized mass spectrometry sample prep options
  • Mass Spectrometry:
    • Customized LC gradient coupled with optimal mass spectrometry scan settings for each project
    • Optional: Advanced deep scan with AcquireX work flow until all ions in the sample are fragmented, maximizing the identification and quantitation of low abundance HCPs
  • Data Analysis:
    • Detection and absolute quantitation of HCPs and DS using BioPharma Finder
    • Comprehensive HCP quantitation analysis, producing reports aligned with regulatory requirements for biologics submissions

HCP identification and quantitation procedure

HCP Identification Procedure

HCP identification and quantitation experimental procedure:

  • HCP sample preparation: Optimized for different HCP sample types; covers cleaning up sample & concentrating (if needed) followed by measuring protein concentration
  • Optional protein fractionation: Fractionate proteins to separate HCPs from DS
  • Trypsin digestion: Reduction, alkylation and trypsin digestion
  • LC-MS/MS with optional AcquireX work flow: Digested peptides are separated and analyzed by Orbitrap Exploris high-resolution mass spectrometer
  • Data analysis by BioPharma Finder: Data contains protein ID and quantitation, as well as peptide level details

Sample Info

Please follow these general principles regarding biohazardous materials and buffer condition in getting your samples ready.

Protein amount for each gel: 20-50 µg of total protein from each sample.

Protein concentration: 5-20 mg/mL

Buffer: Please feel free to submit samples in whichever buffer that you would normally use. The sample preparation including protein extraction (if applicable), sample cleaning and measuring protein concentration is not covered by the LC-MS/MS cost. Additional charge may apply on samples with concentration less than 4 mg/mL, or require significant amount of the extra work such as protein eluting, sample pooling, extensive buffer exchange or serum abundant protein depletion.

Sample type: Please inform our scientists about your sample type. We will send you additional tips for each sample type.

Pricing

Services DescriptionAcademic/Governmental LabsIndustrial/Commercial Labs
CodePriceCodePrice
HCP Identification & Quantitation360Acall for price360Ncall for price
Protein Fractionation355Acall for price355Ncall for price
AcquireX Workflow356Acall for price356Ncall for price

Turnaround time is ~9 days. Price covers the following:

  • Experimental design
  • Reduction, alkylation and Trypsin digestion
  • Protein fractionation – optional
  • LCMSMS with optional AcquireX work flow
  • Standard database search against HCP/DS database using BioPharma Finder
  • Data report