Commercial HCP-ELISA

Determine the total HCP quantity in a sample
Optimize your HCP ELISA protocol
Validate the ELISA's coverage of your drug and process

Technical details, sample info, experts etc.

Measure HCP level with a generic assay or process-specific ELISA antibodies

Host cells used for recombinant protein expressions such as CHO, E. coli, and HEK293 contain thousands of proteins (HCPs) that contaminate the purified protein. An HCP assay using a commercial ELISA enables a quick assessment of the HCP amount. It is suitable for both process samples and the final biopharmaceutical product.

HCP impurities ...

Measure HCP level with a generic assay or process-specific ELISA antibodies

HCP impurities may affect drug safety, and they are crucial to remove from your biologic at an early stage. As stated in USP 39 (US Pharmacopeia Chapter 39, 2016), “HCPs should be reduced to low levels.”

A wide range of commercial HCP assay is available. We offer to analyze your samples by any of them, e.g., HEK293, CHO, E. coli, etc. We can also optimize the ELISA protocol for your samples – and provide validation for your process by HCP ELISA coverage analysis.

Which HCP assay service should you choose?

As standard, you can select between the two services described below:

Option 1: HCP-ELISA analysis of your samples

Use of either a commercial kit (e.g., HEK293 or CHO) or process-specific ELISA antibodies.
You get an analysis of your process samples or final product.
Receive a report with the ELISA HCP concentration by email.

Option 2: Optimize and validate your HCP-ELISA

We can provide a coverage analysis of any commercially available HCP assay or your process-specific antibodies.

First, we optimize the ELISA antibody-HCP binding
Next, the HCPs are immunocaptured with the ELISA antibodies
Then, the total and affinity-purified HCPs are identified by SWATH mass spectrometry
Finally, you get a report with the HCP coverage percentage and a list of immunocaptured and total HCPs

Send us a request, and we will adjust the analysis to meet your specific needs.

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"Using the Alphalyse LC-MS/MS coverage method in HCP ELISA selection, we estimate a savings of approximately $1M and, likely, one year of development time"

SAVARA Aps, Horsholm, Denmark Lars Skriver, Senior Science Officer

"The most sensitive, yet robust, Host Cell Protein analysis we’ve seen"

Analyses of key protein features, stability and impurity levels

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We would like to help you as much as possible with your project and therefore provide several kinds of customer support:

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Technical details

Introduction to ELISA

Enzyme-Linked Immunosorbent Assay (ELISA) is an immunoassay that detects the presence of a specific ligand. The ligand is often a protein, which is then recognized by the assay’s antibodies. Thus, developing a kit of antibodies from a particular pool of ligand makes it possible to assess the presence and approximate amount of target proteins quickly.

Therefore, an HCP assay using ELISA is commonly used for performing HCP analysis of biopharmaceuticals. Examples include therapeutic proteins, recombinant peptides, vaccines, monoclonal antibodies (mAbs), antibody-drug-conjugates (ADCs), biosimilars, and gene therapy products.

Sandwich ELISA for host cell protein analysis

Steps of a sandwich ELISA: 1) We coat Anti-HCP capture antibodies to the ELISA plate. 2) HCP antigens are added to wells and bind to corresponding antibodies. 3) Detection antibodies are added and bind to antigens. 4) Enzyme conjugate is added and binds to detection antibodies. 5) Finally, a substrate is added, and colorimetric analysis estimates total HCP concentration.

Despite their widespread use, sandwich ELISAs rely on antibody specificity and rarely cover all HCP species in a process step or final product. It is thus essential to validate all ELISA antibodies before using them for Host Cell Protein detection. For this purpose, an HCP ELISA coverage analysis is performed, e.g., by ELISA-MS.

Alphalyse services

Our setup is suitable for all HCP ELISAs, including those targeting the expression systems:

Mammalian: CHO, HEK293, HeLa, NS0, SP2/0
Bacteria: E. coli, L. lactis, P. fluorescens, S. aureus
Yeast: S. cerevisiae, P. pastoris

It is also suitable for individual process reagents, such as human serum albumin (HSA), bovine serum albumin (BSA), and Protein A. Moreover, we work with commercial/generic, process-specific, and platform assays from common HCP kit manufacturers like Cygnus (HEK293), Rockland, Biogenes, etc.

Sample preparation

An HCP assay requires both ELISA antibodies and a mock cell lysate or early process sample.

Antibodies
1. The ELISA optimization and immunocapture analysis requires typically a minimum of 100 ug of antibodies.
2. The samples should not contain large amounts of detergents
3. The concentration should be >0.5 µg/µl
4. Use Eppendorf Safe Lock tubes or similar high-quality tubes from your lab
5. Freeze or refrigerate to +4 ^oC for cold shipment of the liquid sample
MOCK/early process sample
1. The mock/early process sample requires typically a minimum of 100 µg of protein material
2. The samples should not contain large amounts of detergents
3. The concentration should be >0.5 µg/µl
4. Use Eppendorf Safe Lock tubes or similar high-quality tubes from your lab
5. Freeze or refrigerate to +4 ^oC for cold shipment of the liquid sample

Please contact us for more specific sample requirements before shipment.

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