Epitope Mapping

Overlapping peptide libraries, peptide pools/subpools, and mapping panels for antibody and T-cell epitope identification — with defined specs and documented QC. Capabilities: mg–gram scale per peptide, crude/≥95%/≥98% purity tiers, and optional HPLC/UPLC + LC‑MS verification (sequence‑dependent).

Overview

Epitope mapping identifies which region(s) of an antigen are recognized by an antibody or immune receptor. It is commonly used to characterize specificity, localize binding regions, and guide follow-on work such as antibody engineering, assay development, or vaccine research [1].

A practical starting point is distinguishing linear epitopes (continuous sequence segments) from conformational epitopes (residues brought together by protein folding). Overlapping peptide libraries and follow-on scan panels are especially useful for localizing linear epitopes and refining minimal motifs [4].

For epitope selection context and standardized identifiers, researchers often reference the Immune Epitope Database (IEDB) and NIAID’s IEDB overview [1], [2].

High-throughput peptide synthesis and peptide library workflows supporting epitope mapping projects

Related pages: Peptide pools, Overlapping peptide libraries, and Vaccine peptides.

Antibody epitope mapping T-cell epitope mapping Linear epitope mapping Overlapping peptides Scan panels (alanine / variants) Pools / subpools Variant panels Alanine scan

Use-cases we support

Antibody epitope mapping (linear)

Overlapping peptides to localize binding regions, then truncations/scan panels to define minimal motifs.

Epitope binning & specificity checks

Variant panels and comparative sets to assess cross-reactivity and binding sensitivity.

Vaccine & immunology workflows

Overlapping peptide sets and pools/subpools for screening and deconvolution workflows.

T-cell epitope mapping inputs

Fixed-length overlapping libraries and curated pools aligned to ELISpot/ICS readouts.

Mutation/variant coverage panels

WT vs variant mapping panels to compare binding or immunogenicity in strain/variant contexts.

Custom controls & method development

Positive/negative controls, spacer/linker options, and case-by-case formatting for downstream platforms.

Specs & scale

Mapping projects rank well when they are explicit about specs, QC packages, and scale tiers. Use the table below as a clear “what we can deliver” summary for quote-ready programs.

Capability Options / notes Typical use
Peptide length Common mapping libraries use fixed lengths (e.g., 12–20 aa) plus follow-on truncation panels. Localize binding regions; refine minimal motifs
Overlap scheme Example: 15-mers with 10-aa overlap (5-aa step). Alternative step sizes available. Resolution vs library size tradeoff
Purity tiers Crude, ≥95%, ≥98% (or project-defined target). Screening vs confirmatory panels
Scale From mg-scale per peptide to multi-gram and large-scale resupply (sequence-dependent). Repeat studies; expanded programs
Optional QC Analytical HPLC/UPLC purity profile + LC-MS intact mass (when feasible); documentation per project. Release criteria alignment and traceability
Sequence inputs we accept
  • FASTA, CSV, or Excel peptide lists
  • Region coordinates / protein positions (if applicable)
  • Overlap scheme and library size targets
  • Variant/control definition (WT vs variant, truncations, scans)

If you’re exporting from IEDB, include IDs or exported lists to reduce reformatting [1].

Best keyword focus (for this page)
  • Epitope mapping services (primary)
  • Antibody epitope mapping
  • Overlapping peptide library
  • Peptide pool epitope mapping
  • Linear epitope mapping

Library Design Options

Library format strongly impacts resolution, total peptide count, and downstream screening efficiency. In addition to custom manual design, Bio-Synthesis provides peptide library design software to assist with overlap schemes, truncation panels, and screening layouts. View Peptide Screening Tools.

Peptide Library and Array Designs including overlapping peptides, truncation panels, alanine scanning, T-cell peptides, PTM arrays, and mutation panels

Common peptide library strategies: overlapping libraries, truncation panels, alanine scanning, PTM arrays, T-cell peptides, and mutation panels.

Overlapping Peptide Libraries

Fixed-length peptides tiled across a target region (e.g., 15-mers with 10-aa overlap) to localize linear epitopes and define binding regions.

Explore Overlapping Peptide Library Design→

Truncation Panels

Systematic N- and C-terminal truncations to refine minimal epitope motifs following initial library screening.

Learn more about Truncation Panels →

Alanine Scanning

Single-residue substitutions (commonly alanine) to identify key binding determinants and critical contact residues.

View Alanine Scanning Methods →

T-Cell Peptide Libraries

Class I and Class II compatible peptide sets formatted for ELISpot/ICS and cellular immune screening workflows.

Learn more about T-cell Peptide Libraries →

PTM Arrays

Parallel panels comparing unmodified vs post-translationally modified (PTM) peptides to assess PTM-dependent binding.

See PTM Peptide Arrays →

Mutation / Variant Panels

Wild-type vs variant peptides to evaluate cross-reactivity, strain differences, and mutation-driven binding changes.

Learn more about Mutation Panels →

Peptide pool composition & verification

Pre-pooling checks

Project-defined confirmation steps can be applied before pooling (e.g., LC-MS/HPLC checks where feasible).

Cross-contamination controls

Defined lists, traceable handling, and pool/subpool design choices help reduce ambiguity and support reproducibility.

Optional marker verification

Marker peptide confirmation can be discussed for select pools based on pool size, assay sensitivity, and acceptance criteria.

Pool QC approaches depend on pool size, peptide properties, and intended assay. Include your pool format and acceptance criteria in the quote request.

Practical workflow

1) Define target & assay

Send sequence/region and readout (binding/ELISA/ELISpot/ICS) plus desired resolution.

2) Design library

Pick length/overlap, add controls, and define variants/truncations as needed.

3) Screen and refine

Screen initial library (individual or pooled), then refine with scan/truncation panels.

Quality control & deliverables

Analytical QC

Analytical HPLC/UPLC purity profile and LC-MS intact mass confirmation (when feasible).

Documentation

CoA and project-defined documentation (sequence list, pool composition record, naming conventions).

Resupply readiness

Traceable lists and consistent formatting help repeat orders match confirmed epitope reagents.

FAQ

What purity do I need for mapping libraries?

Screening libraries often use crude or ≥95% depending on assay sensitivity and peptide properties. Confirmatory panels commonly specify ≥95% or ≥98%. Define acceptance criteria in your quote request.

What is an overlapping peptide library?

A set of peptides that tile across a protein/region with a defined overlap to localize binding regions and support refinement to minimal motifs.

How do I design 15-mers with 10-aa overlap?

A common approach uses a 5-aa step (15 length minus 10 overlap). Provide the region and we can format the library list.

Can you help with epitope selection inputs?

Yes. If available, include IEDB IDs/exports and any strain/variant context. [1][2]

Do you support pools and subpools?

Yes. Pools reduce handling; subpools help localize signal and simplify deconvolution. Pool verification options are available case-by-case.

Can you include PTMs or other modifications for mapping?

Yes. PTM arrays (unmodified vs modified) and other project-defined modifications can be incorporated when you need to evaluate modification-dependent binding or method development.

How should I choose peptide length and overlap?

Choose based on assay and resolution needs. A common scheme is 15-mers with 10-aa overlap (5-aa step), then follow-on truncations or fine-step panels to define minimal motifs.

Can you map conformational epitopes?

Conformational epitopes may require structure-informed approaches. Constrained/cyclic peptide designs or targeted panels can be discussed depending on the project.

What is a peptide pool, and when should I use subpools?

A peptide pool combines multiple peptides into a single mixture for efficient screening. Subpools split a large pool into smaller sets so you can localize signal faster before testing individual peptides.

What quantities are typical for epitope mapping programs?

Initial screening libraries are commonly supplied at mg-scale per peptide. Confirmatory peptides and resupply lots can be scaled as needed (sequence-dependent) once targets are prioritized.

Request a Quote

What to provide
  • Target sequence/region (FASTA) and any known epitope region
  • Peptide length + overlap (or request a recommendation)
  • Library size, variants/controls (WT vs variant, truncations, scans)
  • Purity target(s) + quantity per peptide
  • Intended assay (binding / ELISpot / ICS / ELISA)

Share your design details and timeline. We’ll recommend a feasible library plan and an appropriate analytical characterization package.

Fastest path

Recommended Reading

  1. [1] Vita, R. et al. The Immune Epitope Database (IEDB): 2018 update. Nucleic Acids Research (2019). DOI
  2. [2] NIAID (NIH). Immune Epitope Database and Analysis Resource (IEDB). Resource overview
  3. [3] PEPperPRINT. PEPperMAP® services / peptide microarray epitope identification. Service page
  4. [4] IEDB.org. IEDB database and analysis tools. iedb.org

Links are provided for scientific background; access may depend on institutional subscriptions.

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