Antibody Sequencing 2018-04-16T09:57:36+00:00

Antibody Sequencing

What Is Antibody Protein Sequencing?

Antibody protein de novo sequencing technology can determine the amino acid sequence of an unknown antibody with mass spectrometry. Antibody protein sequencing does not require access to the cell line or prior knowledge of the DNA sequence. The technology is based on mass spectrometry measurements of the protein molecules. Merely 0.2mg of the antibody protein is required. Within weeks you can receive a detailed report, which includes the amino acid sequences of the heavy and light chain antibody sequences, the supporting data for the correctness of the antibody protein sequence, as well as other observations made about the molecule.

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Antibody Sequencing Services

Key Benefits of Antibody De Novo Sequencing

  • Antibody resurrection:  Lost hybridoma? No problem. You can reproduce the antibody recombinantly from 0.2mg or more of mAb proteins. (Talk to us if you do not have 0.2mg.)
  • End-to-end antibody sequencing: optimized high-resolution mass spectrometry workflow and proprietary antibody de novo sequencing software ensure that the whole antibody protein is sequenced from n-term to c-term. No amino acid is left behind.
  • Unparalleled accuracy: The identity of each amino acid in the sequence is supported by strong signal peaks in multiple MS/MS spectra of distinct peptides.
  • Wide applicability: the technology works for all antibody types (IgG, IgA, IgM, IgY, Fab, ScFv, etc.) and species (human, mouse, rat, rabbit, hamster, etc). We have also frequently sequenced conjugated and contaminated proteins, as well antibodies with two different light chains.
  • Dedication to Perfection: Our business is dedicated to mass spec protein sequencing. We have optimized our workflow and software ruthlessly to sequence even the toughest antibodies and to offer the service with the highest quality.
  • See the unseen: Antibody de novo Sequencing frequently identifies unexpected sequence variants and irregular glycosylation sites on the antibodies, which cannot be seen by DNA sequencing.
  • High throughput: This approach works whether you have only one antibody or hundreds of antibodies to sequence

Sample Requirements for Antibody Sequencing

Typical Sample Requirements: monoclonal antibody, quantity>0.2mg, purity>95%.

Deliverable: A comprehensive report including the full sequences of the heavy and light chains, the supporting data for the sequence correctness, and other observations such as irregular modifications on the antibody. (Click here for and example of a coverage report.)

Turnaround time: Typically 1-2 weeks after receiving the sample.

Other Antibody Sequencing Techniques

Hybridoma Sequencing

Hybridoma sequencing sequences the cDNA encoding of the variable heavy (VH) and variable light (VL) domains of the hybridoma cell line. This is a mature technology and the most economical method when the antibody secreting cell line is available.

Edman Sequencing

Edman sequencing has been around for several decades. It directly sequences the protein, but usually only capable of sequencing peptides up to 50 amino acids long. It is a low throughput process.

Intact Mass Analysis and Peptide Mapping

Intact mass analysis and peptide mapping are very often mistakenly associated with the sequencing technology. In fact those are great confirmation tools when the sequences of the antibody proteins are known. Intact mass analysis can confirm at the protein level whether the molecular weight is the same as the experimental observation. Peptide mapping can confirm at the peptide level, that the precursor of the peptides are observed. Both technologies lack the insights at the amino acid level, which is absolutely required in sequencing.

Antibody Sequencing

Also known as de novo antibody sequencing, this is direct sequencing of the antibody’s amino acid sequence using tandem mass spectrometry without reference to a protein or antibody sequence database. For more information, see Peptide Mapping vs Antibody Sequencing

Try REmAb™ Antibody Sequencing Service
Special offer 40% off your first order:

Antibody Sequencing Services