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Edman sequencing of mAb

N-terminal sequence analysis of monoclonal antibodies

Do you need to know the amino-terminals of the heavy and light chains of your mAb?

We can sequence up to 30 residues from the N-termini of your antibody by N-terminal Edman degradation. Our experts perform N-terminal antibody sequencing, using two Procise 494 Edman sequencers.

You obtain the most accurate sequences on purified heavy chains and light chains. Typically, we reduce the antibody and separate the HC and LC by 1D SDS PAGE. The gel is then electroblotted onto a PVDF membrane. We now load the stained PVDF bands of the HC and LC onto to the Edman sequencer. Edman degradation is a cyclic procedure, which cleaves off one amino acid at a time. Afterwards it is identified by chromatography, and 10 cycles are sufficient to determine the N-terminus of most samples.

Antibodies are often modified in the N-terminals by partial formation of pyroglutamic acid. This blocks the terminal and thus prevents the Edman sequencing chemistry from working properly.  We can remove the pyroglutamic acid by enzymatic treatment to enable the Edman sequencing.

Our N-terminal antibody sequencing service includes:

  • First step is 1D SDS PAGE separation of the HC & LC.
  • This is followed by blotting onto PVDF membrane.
  • Next step is staining and cutting of the HC & LC bands.
  • Then we perform Edman sequencing up to 30 amino acids.
  • If needed, we can add de-blocking of pyroglutamic acid (optional).
  • We deliver the sequencing report with easy to understand results, which you can use for your studies.

Download application note (pdf – 1 MB) Learn about the many applications of Edman degradation. – Then apply them to your studies for improved results.

Looking for specific residues? Then send us a request for a customized project.

 

Services/Products
Protein analysis by Alphalyse

N-terminal Edman sequencing antibody chains, 1D PAGE & PVDF blot

  • 1D PAGE separation of antibody HC & LC, PVDF blot, staining & cut of 2 bands
  • Edman sequencing of HC & LC, 10 residues
  • Comparison with customer sequences

$2,650.00

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote
Protein analysis by Alphalyse

N-terminal Edman sequencing antibody chains – deblocking pyroGlu

  • 1D PAGE separation of antibody HC & LC, PVDF blot, staining & cut of 2 bands
  • PyroGlu removal by enzyme, Edman sequencing 10 residues
  • Comparison to customer sequences

$3,350.00

World Class Service
Quality analysis, presented in easy-to-understand reports
Info / Order
Request quote

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.

  • 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
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.
  • 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.

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. 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!

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

Order analysis now
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