Custom Antibody Production

Overview — Custom antibody production for proteinase research

Custom antibody production is the process of raising an antibody against a target of your choosing, in the host species that best suits your downstream application, with the immunogen designed to give you the specificity your experiment requires. Triple Point Biologics has been running custom polyclonal antibody projects for 32 years, and our work is specifically tuned for the proteinase family — the metalloproteinases, cathepsins, ADAMs, ADAMTSs, granzymes, kallikreins, serpins, and other proteases whose closely-related paralogues make antibody design non-trivial.

The standard workflow begins with antigen design or antigen acceptance, proceeds through a host-appropriate immunisation schedule (typically 10–14 weeks; rabbit standard is 12 weeks) at a qualified animal facility, and concludes with antigen-affinity purification against the original immunogen. Typical yields depend on host species: 5–15 mg per rabbit, 20–50 mg IgY per chicken, 100–300 mg per goat or sheep, and 1–5 mg per mouse. Every project is delivered with a Certificate of Analysis that includes the immunogen sequence, ELISA titer curve, and Western blot validation data on the final purified material.

What distinguishes this custom antibody production service is the dual-format option. Rather than running a separate hybridoma or phage-display campaign to obtain a sequence-defined reagent, researchers can elect recombinant conversion at project initiation or after reviewing polyclonal performance. The variable regions are cloned from the immunised animal and expressed in CHO cells, producing a recombinant IgG with identical epitope specificity and consistent lot-to-lot supply — critical for validated targets expected to require antibody across multiple experimental years or across multiple collaborating labs.

For researchers studying proteinase families, splice variants, or closely related paralogues, TPB's epitope design consultation draws on three decades of mapping domain-specific sites. Where a pan-family reagent would create signal ambiguity, a well-placed epitope in a divergent loop or non-catalytic domain can resolve the problem before the first injection. This consultation is included as part of the antigen design phase for eligible projects — the same expertise that produces our Superpooled Antibody Kits is applied to your custom project.

When to choose custom antibody production vs. catalog

Custom antibody production makes sense when the reagent you need does not exist commercially — or where existing commercial antibodies have failed in your system, don't discriminate between paralogues you need to distinguish, or have documented lot-to-lot variability that has compromised your project. It is the appropriate route when:

By contrast, if your target is well-represented in a specialty catalog like TPB's own 981-antibody catalog, the off-the-shelf reagent is almost always the correct choice — same 32-year expertise, higher validation depth, and no immunisation campaign required.

Host species selection — a comparison

Host species choice has substantive downstream consequences. Yield, isotype, cross-reactivity considerations, and application compatibility all vary with host. The recommended host depends on the intended use case:

Immunogen design — where custom projects succeed or fail

The single most important decision in custom antibody production is immunogen selection. A poorly-chosen immunogen produces low-titre antibody, cross-reactive antibody, or antibody that fails in your intended application no matter how good the immunisation and purification workflow is. Immunogen selection is where TPB's 32 years of proteinase-family expertise concentrates.

Peptide immunogen design principles

Peptide immunogens (15–20 amino acids, coupled to KLH carrier) are TPB's default recommendation for most projects. Peptide immunogens allow precise epitope control, avoid cross-reactivity with unrelated proteins that share partial homology, and eliminate the batch-to-batch variability of recombinant protein immunogens. Key design considerations:

• Sequence uniqueness: BLAST the candidate peptide against paralogues and orthologues. A peptide with less than 60% identity to the closest paralogue is likely to give paralogue-selective detection. Sequence identity higher than 80% will typically produce cross-reactive antibodies.
• Predicted antigenicity: Peptides in solvent-exposed regions (hydrophilicity + flexibility) produce higher-titre antibodies. Buried regions in the mature fold rarely make good immunogens.
• Post-translational modification sites: Peptides containing or immediately flanking phosphorylation sites, glycosylation sites, or ubiquitination sites can produce antibodies whose signal is masked when the modification is present. Avoid unless PTM detection is the goal.
• Cleavage site avoidance: Peptides spanning known cleavage sites can produce antibodies that only detect uncleaved forms of the target. For proteinases especially, avoid peptides that cross known autoactivation, cathepsin, or other regulated cleavage sites unless cleavage-specific detection is the design goal.
• Domain-specific placement: For a multi-domain target, choosing which domain to peptide-map determines what functional state your antibody detects. Propeptide peptides detect zymogen; catalytic-domain peptides detect total target; C-terminal peptides distinguish shed from anchored forms.

These are the same considerations used to design the domain-specific antibodies in the Superpooled Antibody Kit method. Every custom project at TPB gets a formal antigen design memo — a written record of why the specific peptide was chosen, what paralogues it distinguishes, and what functional forms it is expected to detect.

Recombinant protein immunogen design

Recombinant protein immunogens (full-length or domain-specific) are the alternative when a peptide will not work — typically for antigens that require a folded three-dimensional epitope, where the target has no linear-peptide epitopes with good properties, or where the customer already has purified recombinant protein available. TPB accepts customer-supplied recombinant protein antigens if quality-controlled for purity and concentration.

Recombinant protein immunogens produce polyclonals with a broader epitope repertoire — good for maximum sensitivity and application robustness, less good for paralogue selectivity. Where selectivity is required, peptide is the better choice.

Recombinant conversion — the reproducibility unlock

Standard polyclonal antibodies are inherently lot-variable. Every immunisation campaign generates a slightly different antibody population; titre distribution shifts between bleeds; minor cross-reactivities can emerge or disappear across lots. For a research project running over 2–5 years, or for a reagent needed across multiple collaborating laboratories, this variability creates real reproducibility risk. Recombinant conversion solves it.

The process: after the polyclonal antiserum is validated in the customer's application, B-cells from the immunised animal are collected, the variable region sequences from the antigen-specific clones are amplified by PCR, and the variable regions are cloned into a CHO expression construct. The resulting recombinant IgG has identical epitope specificity to the polyclonal that was validated — because it comes from the same B-cell repertoire — but is now a sequence-defined reagent produced by transfection of a stable cell line.

Detailed workflow

1. Antigen design & acceptance (Weeks 1–2): Peptide immunogen design by TPB (or review of customer-supplied antigen/sequence). BLAST paralogue screening and antigenicity prediction. Custom peptides synthesised by a qualified vendor if required. Epitope mapping consultation included for isoform-targeting projects. Written antigen design memo delivered to customer for approval before proceeding.
2. Immunisation (Weeks 3–14): Host-appropriate immunisation protocol at a qualified animal facility (typically 10–14 weeks; rabbit standard is 12 weeks). Boosts scheduled per protocol; interim bleed titre monitored by ELISA against immunogen after week 6 and week 10.
3. Terminal collection & titre check (Weeks 15–16): Terminal bleed collected; crude serum titre confirmed by ELISA against immunogen before proceeding to purification. If terminal titre is below target threshold, additional boost cycle or alternate animal recommendation made.
4. Antigen-affinity purification (Weeks 17–18): Purified polyclonal antibody by antigen-affinity chromatography (Protein A/G purification available on request). Purified material subjected to endotoxin screening and concentration determination.
5. Optional — Recombinant conversion (Weeks 19–24): B-cell collection from immunised animal; variable regions amplified and cloned; transient CHO transfection for initial expression; stable CHO line generation if long-term supply is anticipated. Sequenced expression construct provided to customer.
6. QC & delivery (Final week): Western blot validation run on final purified material against relevant positive-control lysate or recombinant protein. ELISA titre confirmation. CoA compiled with immunogen sequence, ELISA titre curve, WB validation image, purification method, concentration, storage buffer, and QC date. Antibody shipped on dry ice with storage recommendations.

Polyclonal-only total: approximately 18 weeks. With recombinant conversion: approximately 24 weeks.

Deliverables

• Purified polyclonal antibody (antigen-affinity or Protein A/G; typical yield depends on host: 5–15 mg per rabbit, 20–50 mg IgY per chicken, 100–300 mg per goat or sheep, 1–5 mg per mouse)
• Certificate of Analysis (CoA) including:
  • Immunogen sequence or peptide identity
  • Purification method and column details
  • ELISA titre curve against immunogen
  • Western blot and/or ELISA validation data on final lot
  • Protein concentration and storage buffer composition
  • Endotoxin level (if requested)
  • Purification date and lot number
• Storage and handling recommendations (buffer, aliquoting guidance, recommended temperature, freeze-thaw tolerance)
• Antigen design memo — written record of why the specific immunogen was chosen and what functional form it should detect
• Recombinant conversion adds:
  • Sequenced variable-region expression construct (delivered as annotated plasmid map)
  • CHO-expressed recombinant IgG lot with matching CoA
  • Stable CHO cell line (if long-term supply is anticipated)

Options

• Antigen type: TPB-synthesised peptide immunogen (via qualified vendor) or customer-supplied peptide / recombinant protein antigen
• Purification method: Antigen-affinity purification (standard) or Protein A/G polyclonal purification (optional)
• Output format: Polyclonal IgG only (~18 weeks) or Polyclonal IgG + recombinant conversion (~24 weeks)
• Epitope design consultation: Domain-specific epitope design for researchers targeting specific isoforms or paralogues (included in antigen design phase for eligible projects)
• Species reactivity screening: Cross-reactivity against related paralogues or ortholog species tested at QC as part of the final validation (available on request)
• Conjugate variants: Biotin, HRP, or fluorophore conjugation of the final purified antibody available as an add-on

Timing and pricing

Polyclonal-only: approximately 18 weeks from project initiation to delivery.
Polyclonal + recombinant conversion: approximately 24 weeks.
Pricing: project-based quote following a brief antigen/project review. Contact TPB with your target, preferred antigen format, and output requirements to receive a written quote within 2 business days.

Pricing is not published as a list because project scope varies substantially depending on antigen complexity, host species, animal count required, whether recombinant conversion is included, and any custom QC requirements. Every project is quoted individually to ensure the quote matches what will actually be delivered.

Quality standards and QC

TPB custom antibody projects are run to standards developed for our own catalog production — the same 981-antibody catalog that has been in continuous production since 1994. Every animal is immunised at a qualified veterinary-supervised facility; every purification uses HPLC-controlled columns; every final lot receives Western blot validation before delivery. If a lot fails QC — insufficient titre, wrong band size, high background — the project is either re-immunised at TPB's expense or refunded per the project agreement.

For projects requiring documentation for regulatory or institutional audit purposes, TPB provides audit packets on request containing production records, animal facility credentials, purification traces, and QC data. See our quality and validation approach for the standards TPB maintains on catalog production — the same standards apply to custom work.

IP and confidentiality

Every custom project begins with a signed project agreement covering confidentiality, IP ownership, and material transfer. Standard terms: the customer owns the resulting antibody and any recombinant sequence, TPB retains a non-exclusive right to reference the project internally (never publicly without permission), and no third parties are granted rights without customer approval. For projects requiring specific institutional MTAs or novel IP clauses, TPB works with customer legal to reach appropriate terms before work begins.

Scope — what is and isn't offered

• Mouse monoclonal antibody production (hybridoma-based) is not offered
• Phage display and in vitro antibody selection are not offered
• Hybridoma generation is not offered
• Diagnostic or therapeutic reagent production is outside scope — all antibodies are produced for research use only (RUO)
• Clinical or GMP manufacture is not offered

Frequently Asked Questions