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Coverage analysis of HCP ELISA

Host cell protein coverage analysis for validation of ELISA

Do you know the HCPs covered by your ELISA?

With our coverage analysis, we evaluate antibodies of generic and process specific HCP ELISA. The host cell proteins in a mock lysate or an early process sample are immunocaptured using ELISA antibodies and identified by LC-MS/MS. As a result, we provide a list of the HCPs recognized by the ELISA antibodies.

You can use the analysis for:

  • Evaluation and comparison of ELISA reagents to select the most suitable for your product and process.
  • Fit-for-purpose validation of generic commercial ELISA kit and also process specific ELISA.
  • Regulatory documentation.

This is an alternative analysis to spot counting in 2D PAGE gels and Western blots, and provides a much more detailed and accurate information. Our Immunocapture LC-MS analysis will expand your knowledge of the ELISA’s specificity and efficiency, as well as reduce the risk of project delays due to poorly documented ELISA.

Our HCP ELISA 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 list of total HCPs in mock lysate.

Send us a request so we can adjust the analysis to meet your specific needs.

Services
Protein analysis by Alphalyse

HCP ELISA coverage analysis

  • Optimization of antibody-HCP binding.
  • Immunocapture with ELISA polyclonal antibodies.
  • Identification of HCPs by LC-MS/MS.
  • Reporting of HCPs covered by ELISA.

Customized Service
Quality analysis, presented in easy-to-understand reports
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The HCP ELISA coverage analysis is a three step procedure.
Step 1: Optimization of binding of HCP to ELISA antibodies
Step 2: Capture of HCP’s
Step 3: Digest and LC-MS analysis for HCP identification
The LC-MS protein identification analysis is conducted before and after immunocapture to compare the  numbers of HCP bound to ELISA antibodies  with the numbers in the original sample.

Introduction
HCP ELISA coverage analysis using mass spectrometry technology requires both ELISA antibodies and a mock cell lysate or early process sample.

  1. 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 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
  2. 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.

shipping
Ship your protein samples by a courier company of your own choice

Place your samples into micro tubes and pack in a suitable transport box. Remember to print and include the sample submission form.

  • Contact a courier company of your choice (FedEx, UPS, DHL, TNT, World Courier etc.). Use the following sentence to describe the content: “Protein material – Non-toxic, non-hazardous, for research purpose only“.
  • Use cool packs or cooling units if your samples should be stored below 5°C/40°F
  • Ship the samples to the office closest to you (Canadian customers, please ship to Europe):
Customers in the USA
Alphalyse, Inc.
200 Page Mill Road, Suite 100
Palo Alto, CA, 94306
USA
Att: Jette Williams
Tel: (650) 543-3193
Customers outside of USA
Alphalyse A/S
Unsbjergvej 4 D
DK-5220, Odense
DENMARK
Att: Anette Rasmussen
Tel: +45 6310 6500
shipping
Order a prepaid FedEx Air Waybill < 1 lb./ 500g

A prepaid FedEx Air Waybill can be used for fast and easy shipment of samples in packages up to 1 lb./ 500g.
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack in suitable transport box. Remember to print and include the sample submission form.
  • Print the PDF document and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to send your samples stored at room temperature, this option is usually the right choice. If you fill your box with cooling packs, the weight will most often exceed 1 lb./500g.

shipping
Order a prepaid FedEx Air Waybill < 10 lb./5kg (e.g. for packages with cooling packs)

This prepaid FedEx Air Waybill can be used for shipment of samples in packages up to 10 lb./ 5kg, with or without cooling packs (do not use dry ice!).
We will generate a prepaid Air Waybill, prefilled with the correct information, and send it to you by e-mail (PDF-format) within 24 hours.
Then follow these simple instructions:

  • Place your samples into micro tubes. Pack them in a suitable transport box. Remember to print and include the sample submission form.
  • Fill with cooling units/cold packs (do not use dry ice!).
  • Print the PDF and stick it on the transport box or just hand it to your FedEx officer upon pickup.
  • Call to request a pickup from your local FedEx office.

If you plan to fill your transportation box with cooling packs, select this shipping option. If you plan to use dry ice, you cannot use this Waybill!

shipping
Get a quote for shipment by World Courier

The World Courier service can be used for shipment of samples of all types. If you select this option, we will contact you to provide a quotation based on location and shipping conditions. You will receive a prepaid Air Waybill which will be invoiced together with your analysis results. This is usually a good option if FedEx does not service your location.
With the prepaid World Courier service you get:

  • GxP compliant processes to ensure maximal security and traceability
  • Door-to-door solution on a dedicated vehicle
  • Possibility of providing qualified packaging, temperature monitoring and dry ice
  • Shortest possible transit-time through “next flight out”-procedure
  • Customs clearance and help with paperwork
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