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)


2D image of Cy-3 labeled Applied Biomics external protein markers (EPMs)


2D image of Cy-5 labeled Applied Biomics external protein markers (EPMs)

Cy3-EPMs / Cy5-EPMs

Overlay 2D image of Cy-3 and Cy5 labeled Applied Biomics external protein markers (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


2D image of Applied Biomics external protein markers (EPMs)

Cell Lysates

2D image of whole cell lysates

EPMs / Cell Lysates

Overlay 2D image of EPM and cell lysate showing locations of Applied Biomics EPMs relative to HCPs

Example 1: Adjust protein ratios using Applied Biomics EPMs

No EPM Adjustment

2D DIGE overlay image without EPM adjustment

With EPM Adjustment

2D DIGE overlay image with EPM adjustment
SpotsRatios without EPM adjustmentRatios with EPM adjustment

Example 2: Monitor HCP Contaminant / product purity using Applied Biomics EPMs

DeCyder image of a process sample

Detecting HCP comtaninant and adjust product purity with EPM in process samples

Calculate HCP Contaminant / product purity

  1. Use DIGE Western blot information to locate all product protein (and their isoforms) – blue spots
  2. Calculate % HCP Contaminant /purity of Product in purification process
  3. 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

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 DescriptionAcademic/Governmental LabsIndustrial/Commercial Labs
2D DIGE Analytical Gel (2 CyDye labeling)101A2$1225101N2$1500
2D DIGE Analytical Gel (3 CyDye labeling)101A3$1375101N3$1685
HCP Quantitative Analysis202A-HCPcall for price202N-HCPcall for price

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