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Full-length antibody sequencing
Determine de novo amino acid sequences for antibody heavy and light chains.
De novo amino acid sequencing
Protein Sequencing Service
Full-length de novo amino acid sequencing for antibodies and other purified proteins, using high-resolution nano-LC-MS/MS and Orbitrap Ascend mass spectrometry.
Every project begins with a technical review of your sample and research objective.
PlatformOrbitrap Ascend
MethodNano-LC-MS/MS
ReportSequence & coverage evidence
Sample amountAs low as 50 μg
What the service can support
Sequence proteins directly from mass-spectrometry evidence.
De novo sequencing determines a protein's amino acid sequence from detected fragments, without requiring prior knowledge of the complete sequence.
Protein sequencing
Determine amino acid sequences for purified proteins including antigens and enzymes.
Isoleucine/leucine differentiation
Distinguish isoleucine and leucine assignments using supporting fragmentation evidence.
Sequence coverage confirmation
Review peptide-level coverage and confidence evidence across the reported protein sequence.
Research and identification
Use sequence information to support the study of biological function or confirm the identity of a protein product.
The analytical platform
Orbitrap Ascend mass spectrometry
A multi-protease nano-LC-MS/MS workflow produces complementary peptide fragments for de novo sequence assignment and confirmation.
Reduction and alkylation
The protein sample is reduced and alkylated before enzymatic digestion.
Multiple protease digests
Four to eight proteases may be used, with digests analyzed separately and in replicate to improve sequence coverage.
Nano-LC-MS/MS analysis
Digested peptides are desalted, concentrated, separated by nano-flow liquid chromatography, and analyzed by high-resolution mass spectrometry.
Sequence assignment
HCD and EThcD fragmentation data are interpreted with analysis software to assign amino acid sequences and support I/L differentiation.
Preparing your sample
Typical sample requirements
These are the usual starting requirements. Final acceptance is confirmed after the sample information has been reviewed.
- Purity
- More than 85%
- Concentration
- Above 0.2 mg/mL
- Sample amount
- At least 50 μg
- Protein contaminants
- No BSA or other contaminating proteins
Special samples: Identify samples containing BSA, mixed antibodies, or proteins coupled to materials such as microspheres or HRP. These samples may require additional evaluation and effort.
How a project works
A simple path from sample review to report.
Sample informationShare the sample type, quantity, purity, concentration, and sequencing objective.
Quote & confirmationConfirm sample requirements, project price, turnaround, and shipping instructions.
SequencingThe accepted sample proceeds through preparation, LC-MS/MS, and sequence analysis.
Report & supportReceive the report and ask follow-up questions about the reported sequence results.
Typical report contents
Useful evidence, clearly organized.
- Full-length heavy- and light-chain amino acid sequences for antibody projects
- Isotype identification and CDR annotation
- Heavy- and light-chain sequence coverage confirmation
- Isoleucine/leucine differentiation with confidence evidence
- Observed findings such as N-glycosylation sites, when identified
Report contents depend on the sample and agreed project scope. A typical antibody sequencing report presents separate heavy- and light-chain results, coverage evidence, and I/L differentiation tables.
Before you submit
Common questions
Typical requirements are shown below. Final acceptance and timing are confirmed during project review.
What samples can be analyzed?
Antibodies, antigens, enzymes, and other purified protein samples can be evaluated for de novo amino acid sequencing.
How much sample is required?
Typical starting requirements are more than 85% purity, a concentration above 0.2 mg/mL, and at least 50 μg of sample. Samples should not contain BSA or other protein contaminants. Final requirements are confirmed before shipment.
Can you work without a reference sequence?
Yes. De novo sequencing assigns amino acid sequence directly from mass-spectrometry fragmentation evidence, so a complete reference sequence is not required.
What is the turnaround time?
Typical analytical turnaround after sample receipt is 1–2 weeks for one sample and 2–3 weeks for 2–10 samples. The project-specific schedule is confirmed with the quote.
Can special or conjugated samples be submitted?
Potentially. Samples containing BSA, mixed antibodies, microspheres, HRP, or other coupled substances should be identified during quotation because they may require additional evaluation or effort.
What affects the project price?
Pricing can vary with sample quality, sample specificity, and the additional effort required for special or complex samples. A detailed quotation is prepared after the sample information is reviewed.
How should the sample be shipped?
Shipping instructions are provided after project confirmation. Sample safety documentation may be required depending on the material being shipped.
What can the report include?
Depending on the project, the report can include the amino acid sequence, heavy- and light-chain coverage, isotype and CDR annotation, I/L differentiation evidence, and identified sequence features.
Have a protein to sequence?
Tell us what you need to learn.
Include the sample type and count, purity, concentration, available amount, known contaminants or conjugates, and any existing sequence information.
Request a quoteRequest a protein sequencing quote
Tell us about your sample so we can review your project.
Request received
Thank you. Your sample information was submitted successfully.
- Confirmation ID
- Received at (UTC)
