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Polyclonal Antibody Production
Highly Specific Antibodies with High Titers
SVL now offers polyclonal antibody services utilizing allotype-defined rabbits developed by NIAID. These rabbits produce highly specific antibodies with high titers.
When ordering a polyclonal project at SVL, you now have the option to use:
allotype-defined rabbits, New Zealand white rabbits, or a combination of the New Zealand and the Allotype-defined Rabbits to broaden your potential response.
For a detailed description of these unique rabbits, please click the link below:
Allotype Defined Pedigreed Rabbits.pdf
Contact us for details and/or a quotation. We look forward to providing you with a great antibody response.
Spring Valley offers two protocols for maximizing antibody production in terms of levels of
antibody and volume of serum. These protocols represent a basic approach that can be extended and
enhanced using the available options listed below or your custom study design.
Standard Protocol
The standard protocol outlined below can be used with almost any antigen, usually of unknown or weak immunogenicity.
| Day |
Procedure |
| 0 |
Pre-immunization test bleed |
| 0 |
Primary immunization |
| 21 |
Immunogen boost |
| 42 |
Immunogen boost |
| 52 |
Standard bleed |
| (ELISA or Western Blot available) |
| 63 |
Immunogen boost |
| 73 |
Standard bleed |
| (ELISA or Western Blot available) |
Fast Protocol
The fast protocol can be used with almost any antigen, usually of known or strong immunogenicity.
| Day |
Procedure |
| 0 |
Pre-immunization test bleed |
| 0 |
Primary immunization |
| 21 |
Immunogen boost |
| 31 |
Standard bleed |
| (ELISA or Western available) |
| 42 |
Immunogen boost |
| 52 |
Standard bleed |
| (ELISA or Western available) |
Protocol Options
The following options are available for tailoring the protocols to produce the antibodies you need: screening
animals (test bleed) for existing antibodies prior to placement on a study, serum filtration
through 0.45 micron filter, Western blots or ELISA as screening/assessment tools, serum antibody
purification, or your customized study parameters. In addition to these options, any protocol may be
extended to obtain a larger volume of serum or to possibly increase antibody titers.
Standard Monthly Extension Protocol
| Extention Day |
Procedure |
| 1 |
Immunogen boost |
| 12 |
Standard bleed |
| 27 |
Standard bleed (or exsanguination) |
Antigen Preparation
Spring Valley can accept antigen for immunization in any form including, but not limited to, in solution, as a
precipitate, as a cell suspension, as a band in acrylamide gel, or as a blot on nitrocellulose. Freund’s
Complete Adjuvant is used for the initial injection and Freund’s Incomplete Adjuvant is used for
subsequent injections. Other adjuvants are available. The recommended minimum antigen dose per
animal is 100 µg protein or 250 µg peptide. Minimum antigen requirements are listed below for several activities.
Minimum Antigen Requirements
| ELISA (up to 6 serum samples) |
20 ug |
| Western blot (up to 6 serum samples) |
20 ug |
| Fast protocol (total per animal) |
750 ug |
| Standard protocol (total per animal, or 5 mice) |
1 mg |
| Protocol extension (total per animal) |
250 ug |
Antibody Production in Many Animal Species
Antibody production can be conducted in a variety of animals. Although the rabbit is, in most instances, the
ideal animal for polyclonal antibody development, mice, rats, guinea pigs, ferrets, sheep, and goats are
also available. The table below indicates expected serum volume per species per protocol.
Approximate serum volumes (ml) per bleed for multiple species
| |
Rabbit |
Rat |
Guinea pig & Ferret |
Mouse |
Sheep & Goat |
| Test bleed |
5 |
0.25 |
0.5 |
0.2 |
15 |
| Fast protocol |
| Day 0 pre-bleed |
5 |
0.25 |
0.5 |
0.2 |
50 |
| Day 31 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| Day 52 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| Standard protocol |
| Day 0 pre-bleed |
5 |
0.25 |
0.5 |
0.2 |
50 |
| Day 52 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| Day 73 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| Protocol extension |
| Day 12 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| Day 27 standard bleed |
15 |
0.5 |
1 |
0.2 |
400 |
| or |
| Day 27 exsanguination |
60 |
5 |
6 |
0.5 |
1500* |
| Exsanguination |
60 |
5 |
6 |
0.5 |
1500* |
| Plasmapheresis |
- |
- |
- |
- |
1500 |
| * Plasmapheresis performed in lieu of exsanguinations. |
Due to the inherent variability between immunogens and animals, Spring Valley does not guarantee the quality or
quantity of the polyclonal antibody.
Peptide Synthesis and Conjugation
Spring Valley protein chemists can analyze your protein sequence to predict potentially immunogenic peptide
sequences. We strongly recommend you use this service unless you have already done so. There is no
fee for this service if you have your peptide synthesized with us. Peptides are synthesized to
greater than 85% purity by HPLC analysis. Other purity levels, analytical methods and modifications
are available upon request. Conjugation of peptides to KLH is standard, but other carriers (ie. BSA, OVA)
are available. Click here to receive a quotation for the synthesis and conjugation of your peptide.
Please contact us to discuss your particular needs. Let us help you.
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