HCP Contaminant Detection
Detecting and monitoring HCP Contaminant / product purity in the drug purification process is another important application of our HCP assay. Due to the different spot patterns from samples in the purification process, External Protein Markers (EPMs) can be added to the sample and used to adjust the HCP quantitation. Applied Biomics has developed several EPMs at different pH and Mw for you to choose from.
The EPMs can serve as a standard to accurately assess product purity in the following areas:
- Evaluating HCP Contaminant / product purity of therapeutic antibodies, recombinant proteins and vaccines
- Monitoring and optimizing of bioprocess
- Characterizing batch to batch cell lines
- Evaluating storage conditions of biological samples
Example 1: Adjust protein ratios by Applied Biomics EPMs
Example 2: Monitor HCP Contaminant / product purity by Applied Biomics EPMs
Applied Biomics External Protein Markers (EPMs)
Cy3-EPMs

Cy5-EPMs

Cy3-EPMs / Cy5-EPMs

- EPM_1: 76 kDa, pI=6.0, 6.2, 6.6
- EPM_2: 30 kDa, pI=5.4 – 6.0
- EPM_3: 14 kDa, pI=6.8 – 7.2
Locations of Applied Biomics EPMs Relative to HCPs
EPMs

Cell Lysates

EPMs / Cell Lysates

Example 1: Adjust protein ratios using Applied Biomics EPMs
No EPM Adjustment

With EPM Adjustment

Spots | Ratios without EPM adjustment | Ratios with EPM adjustment |
1 | 1.6 | 3.9 |
2 | -6.3 | -2.6 |
3 | -18.6 | -7.6 |
4 | 9.2 | -22.5 |
5 | 6.4 | 15.7 |
6 | -11.8 | -4.8 |
7 | -2.2 | 1.1 |
EPM | -2.4 | 1.0 |
Example 2: Monitor HCP Contaminant / product purity using Applied Biomics EPMs
DeCyder image of a process sample

Calculate HCP Contaminant / product purity
- Use DIGE Western blot information to locate all product protein (and their isoforms) – blue spots
- Calculate % HCP Contaminant /purity of Product in purification process
- Optional: Adjust the product ratios between different purification steps using EPMs – green spots
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 is preferred.
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.
Pricing
Services Description | Academic/Governmental Labs | Industrial/Commercial Labs | ||
---|---|---|---|---|
Code | Price | Code | Price | |
2D DIGE Analytical Gel (2 CyDye labeling) | 101A2 | $1200 | 101N2 | $1500 |
2D DIGE Analytical Gel (3 CyDye labeling) | 101A3 | $1349 | 101N3 | $1649 |
HCP Quantitative Analysis | 202A-HCP | TBD | 202N-HCP | TBD |
Price covers:
- Experimental design
- Standard sample preparation and protein concentration determination
- Fluorescent dye labeling using CyDye
- IEF and SDS-PAGE
- Gel image scan using Typhoon scanner
- Image analysis (ImageQuant)
- Data Report