HCP Assay by commercial 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 generic or process-specific ELISA antibodies

Host cells used for recombinant protein expression, such as CHO, E. coli and HEK 293, contain thousands of proteins (HCPs) that contaminate the purified protein. An HCP assay using ELISA enable fast assessment of the HCP amount in both process samples and the final biopharmaceutical product.

HCP impurities may affect drug safety and they are cruci...

Measure HCP level with generic 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 kits are available. We offer to analyze your samples, and can also optimize the ELISA protocol to your samples – and provide validation for your process by HCP ELISA coverage analysis.

Choose from our HCP assay services:

Option 1: ELISA HCP analysis of your samples

Using a commercial kit (e.g. HEK 293 of CHO) or process specific ELISA antibodies
We analyze your process samples or final product
And email you a report with the ELISA HCP concentration

Option 2: Optimize and validate ELISA by HCP coverage analysis

We can use any commercially available ELISA kit 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.

MoreLess details

Start the collaboration

We offer customized solutions, contact us to discuss your project.

Ask a question or get a quote

Send us a description of your project and we shall be happy to present you with a solution.

Set up a phone meeting

We encourage you to call us or schedule a meeting to discuss your project in detail.

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Direct phone numbers

Testimonials

"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

Knowledge center

More information

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

Download & links

E-book: Comparison of Coverage Methods
Poster: HCP Coverage by Immunocapture and LC-MS/MS
Webinar: Which HCP impurities is your ELISA missing?
Video: The risk of using commercial HCP ELISAs in upscaling of batches
Blog Post: Why do I need a ‘Lucky’ goat for precise ELISA results?
Blog Post: Can I use commercial HCP ELISA kits for my clinical trial application?
Article: Combining ELISA-based immunocapture and mass spectrometry

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. By developing a kit of antibodies from a specific ligand pool, it is possible to quickly assess the presence and approximate amount of target proteins.

Therefore, the HCP assay using ELISA is widely used in biopharmaceutical development and quality control – and the industry standard to perform HCP analysis of biopharmaceuticals, such as 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) Anti-HCP capture antibodies are coated to 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 bind to detection antibodies. 5) Finally, substrate is added and colorimetric analysis is done to estimate 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, a 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, HEK 293, HeLa, NS0, SP2/0
Bacterial: E. coli, L. lactis, P. fluorescens, S. aureus
Yeast: S. cerevisiae, P. pastoris

As well as for individual process reagents, such as human serum albumin (HSA), bovine serum albumin (BSA) and Protein A. Moreover, we are highly experienced in working with commercial/generic, process-specific and platform assays from common HCP kit manufacturers like Cygnus (HEK 293), 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.

Related services

You may also find these other HCP services relevant:

Meet the experts

For this type of analysis our experts include:

Katrine Pilely

PhD in Biomedicine and BSc in Biology

Solveig Beck Nielsen

PhD in Molecular Biology

Rikke Raaen Lund

PhD in Biomedicine

HCP Assay by commercial ELISA

Measure HCP level with generic or process-specific ELISA antibodies

Measure HCP level with generic or process-specific ELISA antibodies

Choose from our HCP assay services:

Option 1: ELISA HCP analysis of your samples

Option 2: Optimize and validate ELISA by HCP coverage analysis

The process:

Learn more:

HCP coverage webinar

ELISA-MS Coverage Method Article

Start the collaboration

Ask a question or get a quote

Set up a phone meeting

Testimonials

"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"

Knowledge center

More information

Introduction to ELISA

Alphalyse services

Antibodies

MOCK/early process sample

You may also find these other HCP services relevant:

Meet the experts