Identify and Quantify HCPs
Our LC-MS/MS based analysis can identify and quantify host cell proteins with high sensitivity detection and accurate quantitation. Our platform uses the Orbitrap Exploris with BioPharma Finder for identification and quantitation of HCP contaminants as low as 1-10 ppm in the product. We have optimized our work flow starting from sample preparation, mass spectrometry scan settings to data analysis, specifically:
- Optimized sample preparation for different sample types
- Protein level: Protein fractionation to separate drug substance (DS) from HCPs
- Peptide level: Advanced deep scan with AcquireX work flow until all ions in the sample are fragmented, maximizing the identification and quantitation of low abundant HCPs
- Data analysis: Identify and quantify HCPs and DS with BioPharma Finder
HCP identification and quantitation 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 AquireX 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
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 standard sample preparation is covered by the DIGE gel cost. Such samples should have protein concentration in the range of 5-20 mg/ml. Additional charge may apply on samples that require extensive amount of the extra work such as protein eluting, concentrating, 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.
|Services Description||Academic/Governmental Labs||Industrial/Commercial Labs|
|HCP Identification & Quantitation||360A||call for price||360N||call for price|
|Protein Fractionation||355A||call for price||355N||call for price|
|AcquireX Workflow||356A||call for price||356N||call for price|
Turnaround time is ~9 days. Price covers the following:
- 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