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Immunization and egg-collection procedures for custom chicken polyclonal antibody production

Our comprehensive array of Thermo Scientific Pierce custom antibody services includes a customizable option for chicken immunization, egg-collection and titer analysis for chicken antibody production, plus several extension possibilities.

Custom Antibody Request

Highlights

  • Custom chicken antibodies—a reliable protocol for immunizing chickens, collecting eggs, testing bleeds, reporting titer results and delivering eggs
  • Track and customize project—easily add additional chickens to the default protocol; track and manage projects online with OpenProject Tool: you decide which animals to continue or terminate (see Extensions below)
  • Full service integration—combine this immunization service with our complete set of services for antigen preparation and antibody purification (see Custom Polyclonal Antibodies pages for details)
2-chicken 80-day protocol

This default immunization schedule for chicken antibody production uses two White Leghorn chickens; additional chickens are easily added to the standard order for a modest cost. Injections are subcutaneous (SQ) as emulsions in Complete Freund's Adjuvant (CFA) or Incomplete Freund's Adjuvant (IFA); alternative adjuvants can be used if requested.

80-day chicken immunization protocol for chicken polyclonal antibody production

Antigen preparation, such as peptide synthesis and/or immunogen conjugation, occurs before Day 0. Injection amounts are given for a conjugated peptide antigen (e.g., KLH) or protein immunogen; MAP-peptide antigens are injected at 0.5mg. Protocol Days are approximate (± 2 days).

Procedure Protocol day Description
Control serum collection Day 0 Pre-immune bleed (~2cmL per chicken)
Primary injection Day 1 Immunize with 0.25 mg antigen in CFA, SQ 4 sites
1st booster Day 14 Boost with 0.10 mg antigen in IFA, SQ 4 sites
Egg collection Day 21 Collect eggs for one week
2nd booster Day 42 Boost with 0.10 mg antigen in IFA, SQ 4 sites
Egg collection Day 48 Collect eggs for one week
Serum collection Day 56 Test-bleed (~1-2 mL per chicken)
ELISA and shipping Day 65 ELISA titration (results available online);
Verify disposition of chickens; decide to continue or terminate
Instruction due date Day 80 If no animal instructions are received, per diem charges will apply

Deliverables (Day 65): 32 vials

  • 2 vials: 2 mL per vial pre-immune sera
  • 2 vials: 1-2 mL test bleed post-inject sera (Day 56)
  • 28 vials: post-inject eggs (2 week egg collection)
Protocol extensions

Several pre-defined (yet customizable) options are available for extending or continuing the chicken immunization and egg-collection schedule. The purchase decision to continue antibody production can be made at any time before scheduled termination. For an additional cost, exsanguination bleeds may be obtained at the time of termination, yielding approximately 10-15mL of additional serum per animal.

One-month extension protocol for chicken antibody production

Procedure Protocol day Description
1st Booster Day 1 Boost with 0.10 mg antigen in IFA, SQ 4 sites
Serum collection Day 14 Bleed (~1-2 mL per chicken)
Egg collection Day 10-17 Collect 5-7 eggs per chicken

Deliverables (per chicken): 8 vials

  • 1 vial: 1-2 mL test bleed post-inject sera
  • 7 vials: post-inject eggs (1 week egg collection)

High-dose, one-month extension protocol for chicken antibody production

Procedure Protocol day Description
1st booster Day 1 Boost with 0.20 or 0.5 mg antigen in IFA, SQ 4 sites
Serum collection Day 14 Bleed (~1-2 mL per chicken)
Egg Collection Day 10-17 Collect 5-7 eggs per chicken

Deliverables (per chicken): 8 vials

  • 1 vial: 1-2 mL test bleed post-inject sera
  • 7 vials: post-inject eggs (1 week egg collection)

Aggressive, one-month extension protocol for chicken antibody production

Procedure Protocol day Description
1st booster Day 1 Boost with 0.50 mg antigen in IFA, SQ 4 sites
2nd booster Day 7 Boost with 0.20 mg antigen in IFA, SQ 4 sites
Serum collection Day 21 Bleed (~1-2 mL per chicken)
Egg Collection Day 18-25 Collect 5-7 eggs per chicken

Deliverables (per chicken): 8 vials

  • 1 vial: 1-2 mL test bleed post-inject sera
  • 7 vials: post-inject eggs (1 week egg collection)

One-month auto-termination extension protocol for chicken antibody production

Procedure Protocol day Description
1st booster Day 1 Boost with 0.10 mg antigen in IFA, SQ 4 sites
Egg collection Day 10-17 Collect 5-7 eggs per chicken
Exsanguination Day 21 Bleed (10-15 mL antiserum per chicken)

Deliverables (per chicken): 9 vials

  • 1 vial: 1-2 mL test aliquot from terminal bleed
  • 1 vial: 10-15 mL terminal bleed (minus 1-2 mL for test aliquot)
  • 7 vials: post-inject eggs (1 week egg collection)

Terminal bleed protocol for chicken antibody production

Procedure Protocol day Description
Exsanguination Day 1 Bleed (10-15 mL antiserum per chicken)

Deliverables (per chicken): 2 vials

  • 1 vial: 1-2 mL test bleed post-inject sera
  • 1 vial: 10-15 mL terminal bleed
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