HCP Coverage Analysis

Evaluation and comparison of ELISAs
Low amount of antibody required - 1 mg
Alternative to 2D Gels & Western Blotting

Technical details, sample info, experts etc.

Do you know how well your ELISA covers the HCPs in your sample?

If you need to evaluate and qualify the antibodies of a generic or process specific HCP ELISA we can help you. We use an advanced MS-based coverage analysis method (ELISA-MS^™) which mimics the conditions of the ELISA, for more reliable and accurate results:

In this analysis, the host cell proteins in a mock lysate or an early process sample are immu...

Do you know how well your ELISA covers the HCPs in your sample?

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 not only the total sum of the residual HCPs, but also a list of the HCPs recognized by the ELISA antibodies.

How does ELISA-MS^™ analysis compare to 2D PAGE and Western blot?
ELISA-MS^™ is a proven method recognized as a better alternative to spot counting in 2D gel electrophoresis and Western blot, because it provides much more detailed and accurate information. An Immunocapture LC-MS analysis (ELISA-MS^™) will thus expand your knowledge of the HCP-ELISA’s specificity and applicability to your biopharmaceutical. It may also reduce the risk of project delays due to poorly documented HCP ELISA.

A standard ELISA-MS^™ 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 or early process sample.

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

MoreLess details

Let's work together

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.

Start with an online meeting

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

Set up meeting

Direct phone numbers

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 about HCP coverage analysis in these materials, webinar, and blog posts:

Technical details

The HCP coverage analysis (ELISA-MS^™) includes:

The first step is ELISA setup for optimal antibody-HCP binding.
Next, a 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. It serves as a measurement of total protein content to identify all HCPs. This step compares 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 ELISA 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 standard expression systems:

Yeast: S. cerevisiae, P. pastoris
Bacteria: 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

The coverage analysis also measures specific process residuals, such as human serum albumin (HSA), bovine serum albumin (BSA), and Protein A.

Sample preparation

Sample requirements for MS-ELISA analysis

HCP 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 require a minimum of 1 mg of antibodies, typically.
2. The samples should not contain large amounts of detergents
3. The concentration should be >0.5 µg/µl
4. Use an Eppendorf Safe Lock tube or similar tubes
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 a minimum of 100 µg of protein material, typically.
2. The samples should not contain large amounts of detergents
3. The concentration should be >0.5 µg/µl
4. Use an 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

Don’t hesitate to get in touch with 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

Explore

Anette Holck Draborg

PhD in Immunology

Explore

Flemming Holm Bergholt

Biomedical Laboratory Scientist

Explore

HCP Coverage Analysis

Do you know how well your ELISA covers the HCPs in your sample?

Do you know how well your ELISA covers the HCPs in your sample?

A standard ELISA-MS^™ HCP coverage analysis includes

The process:

Learn more:

HCP coverage webinar

Method Article