HCP Antibody Coverage Analysis

Evaluation and comparison of ELISA kits
Low antibody usage - 1 mg
Alternative to 2D Gels & Western Blotting

Technical details, sample info, experts etc.

Do you know the HCPs covered by your ELISA?

If you need to evaluate the antibodies of a generic or process specific HCP ELISA we can help you through our MS-based coverage analysis method.

In this analysis, the host cell proteins in a mock lysate or an early process sample are immunocaptured using the ELISA antibodies and then identified by LC-MS/MS. As a result, you get a list of the HCPs recognized ...

Do you know the HCPs covered by your ELISA?

If you need to evaluate the antibodies of a generic or process specific HCP ELISA we can help you through our MS-based coverage analysis method.

This is a better alternative to spot counting in 2D PAGE gel and western blot, because it provides much more detailed and accurate information. An Immunocapture LC-MS analysis will thus expand your knowledge of the HCP-ELISA’s specificity and efficiency. It may also reduce the risk of project delays due to poorly documented HCP ELISA.

A standard HCP coverage analysis includes

ELISA setup for optimal antibody-HCP binding
Immunocapture with ELISA polyclonal antibodies
Identification of both total HCPs and affinity purified HCPs by sensitive LC-MS/MS
Reporting of HCPs covered by antibodies and also a list of total HCPs in the mock lysate

Send us a description of your project and we will be glad to discuss it with you.

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

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Testimonials

"They handled the project professionally and rapidly, and the report that was very well written, clearly explaining the findings."

Targovax ASA, Kristiina Hyvärinen – Director, QC, viral products

"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

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

Learn more in these materials, webinar, and blog posts:

Brochure
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?
Article: Combining ELISA-based immunocapture and mass spectrometry
E-book: Comparison of HCP ELISA Coverage Methods

Technical details

The HCP coverage analysis (ELISA-MS) includes:

First, ELISA setup for optimal antibody-HCP binding.
Next, capture of HCPs (antigens) to ELISA antibodies immobilized in 96-well plates.
Finally, digest and LC-MS analysis for HCP identification.

ELISA HCP coverage analysis illustration

Illustration of the ELISA-MS method

An aliquot of the antigen sample (a mock/null cell lysate or early process sample) is digested under similar conditions in parallel without antibodies and serves as a measurement of total protein content to identify all HCPs. This is done to compare the number of HCPs bound to ELISA antibodies with the number of HCPs in the original sample (total protein sample).

Applications of ELISA-MS

We have performed a wide range of projects evaluating commercial/generic, process-specific and platform assays from common HCP kit manufacturers like Biogenes, Canopy, Cygnus, Creative Diagnostics, Rockland, etc.

The hcp coverage analysis can evaluate all HCP ELISAs, e.g. those based on the common expression systems:

Yeast: S. cerevisiae, P. pastoris
Bacterial: E. coli, L. lactis, P. fluorescens, S. aureus
Insect: S. frugiperda (SF9)
Mammalian: HEK293, CHO, HeLa, MRC5, GS-NS0, Vero cell, SP2/0, BHK, A549, MDCK

Along with specific process residuals, such as human serum albumin (HSA), bovine serum albumin (BSA) and Protein A.

Sample preparation

Sample requirements for MS-ELISA

HCP ELISA coverage analysis using mass spectrometry technology 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 1 mg of antibodies
2. The samples should not contain large amounts of detergents
3. The concentration should be >0.5 µg/µl
4. Use a Eppendorf Safe Lock tube supplied, or similar 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 a Eppendorf Safe Lock tube supplied, or similar tubes from your lab
5. Freeze or refrigerate to +4 ^oC for cold shipment of the liquid sample

Please contact Alphalyse for 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:

Solveig Beck Nielsen

PhD in Molecular Biology

Katrine Pilely

PhD in Biomedicine and BSc in Biology

Rikke Raaen Lund

PhD in Biomedicine

HCP Antibody Coverage Analysis

Do you know the HCPs covered by your ELISA?

Do you know the HCPs covered by your ELISA?

A standard HCP coverage analysis includes

The process:

Learn more:

HCP coverage webinar

Method Article

Start the collaboration

Ask a question or get a quote

Set up a phone meeting

Testimonials

"They handled the project professionally and rapidly, and the report that was very well written, clearly explaining the findings."

Targovax ASA, Kristiina Hyvärinen – Director, QC, viral products

"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

Knowledge center

More information

Learn more in these materials, webinar, and blog posts:

Sample requirements for MS-ELISA

Antibodies

MOCK/early process sample

You may also find these other HCP services relevant:

Meet the experts