|Comparison between monoclonal and polyclonal properties for custom antibodies production|
|Monoclonal antibodies||Polyclonal antibodies|
|Specificity|| One hybridoma = one single antibody type, detecting one single epitope present on the immunogen.|
Avoids cross-reactivity issues with other target proteins.
Possible to select a clone the most efficient in the given application (protein function inhibition, flow cytometry, sandwich ELISA)
| Unpurified serum:|
Pool of antibodies, including non-antigen-specific antibodies, binds different epitopes of the antigen.
Can generate background and cross-reactions with other proteins.
| Affinity purified serum:|
Sample purity = 99.99%
Specific for the antigen targeting all accessible and immunogenic epitopes of the immunogen.
Low background and potential cross-reactions with other proteins.
|Sensitivity|| Sometimes less sensitive because of the high specificity for only one epitope.|
A good selection of the most sensitive monoclonal antibody can allow strongly binding ones to be found.
|Sometimes more sensitive due to the wide variety of different antibodies represented in the serum.|
|Lead time||~6 months||~2-3 months (can be less with our Expressway protocol in 28 days)|
|Cost||3 to 4 times more expensive than the cost of custom pAbs.||3 to 4 times less expensive than the cost of custom mAb.|
|Reproducibility|| Antibodies always identical|
Many lots of identical antibodies can be produced over dozens of years.
Possible to sequence the hybridoma to make recombinant antibodies.
| Different antibodies from batch to batch.|
Require new immunizations with other animals.
|Quantities|| Almost unlimited quantities.|
Production possible over many years, as long as clones are viable.
|Maximum of a few mg / animal.|
|Available species||Rat, mouse||Rat, mouse, rabbit, chicken, guinea pig, sheep, and goat.|
As an experienced antibodies manufacturer, ProteoGenix has successfully raised more than 1500 polyclonal antibodies during the past 13 years.
|Rabbit||>1.1 mg of protein or >2.2 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
|Chicken||>2.5 mg of protein or >2.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
|Guinea Pig||>0.7 mg of protein or >1.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
|Rat||>0.7 mg of protein or >1.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
|Mouse||>0.4 mg of protein or >0.6 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
|Goat & Sheep||>10 mg of protein or >10 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.|
* Protein (checked by SDS-PAGE), purity >90%
Lower amounts of antigen can be accepted, just let us know. We also can use your own immunization protocol.Peptide for immunization has to be conjugated to a carrier – we can also do the conjugation step if needed.Conjugation makes the immunogen more immunogenic by making it physically large enough for the immune system to recognize it. We conjugate all the peptides we synthesize usually using keyole limpet hemocyanin (KLH). Other good carrier proteins are BSA and OVA.We also recommend small proteins (less than 15KD) to be conjugated.
Step 1: Define your package
- Antigen from customer
Antigen PurificationPACK 1
Antigen PurificationPACK 2
- Gene Synthesis
E. Coli Protein Production
WB GUARANTEEPACK 3
- Antigen from customer
XPRESSWAY PROTOCOLPACK 1XS
- Anti-2 Peptides
Antigen PurificationPACK 2.2
More information regarding our custom antibodies production services:
ProteoGenix has more than 14 years of experience in this field with more than 1500 antibodies generated including a large number of rabbit polyclonal antibodies.
Our expertise in biological engineering such as gene and peptide synthesis, recombinant protein production and polyclonal antibodies production services allows us to provide very complete services.
Thank to our long experience on these domains (from cDNA to antibody) we can determine the best strategy for each project depending on the needs of each one of our customers.
We offer a One-Stop solution for your polyclonal antibodies generation project as we are able to cover all the different steps if needed:
• Antigen production (peptide synthesis / recombinant protein production)
• Animal Immunizations – 7 species available: Rabbit, Chicken, Mouse, Rat, Guinea Pig, Sheep and Goat antiserum
• Antibodies Purification against protein A or G or against the antigen
• Antibodies Conjugation
As an experienced antibody manufacturer, we have a high quality commitment and that’s the reason why we guarantee:
• ELISA results : 1/32 000 for crude serum and 1/64 000 for purified antibodies (for anti-protein polyclonal antibodies services)
• The final quantity of antiserum or of purified antibodies (depends on the species and the package the amount of rabbit antiserum is higher than guinea pig antiserum but lower than goat antiserum)
• WB guarantee for the anti-protein polyclonal antibodies package n°3
We also offer a phospho-specific polyclonal antibodies package.
Experienced antibodies manufacturer:
For polyclonal antibody development, we have carried out thousands of different projects, in particular for:
Pharmaceutical / diagnostic / veterinary / cropscience / food agribusiness multinationals such as Bayer, Sanofi, Servier, or Virbac.
Public research centers such as CNRS, INSERM and Institut Pasteur in France.
Animals immunization protocol
Our standard immunization protocol is 51 days – 4 injections for anti-protein polyclonal antibody (with possibility to extend the protocol if the guarantees are not reached)
For anti peptide polyclonal antibodies, our standard protocol is 70 days – 5 injections (with possibility to extend the protocol if the guarantees are not reached)
It is usually recommended to immunize animals with a lower quantity of antigen and for a longer time than to immunize the animals on a shorter timing with a larger quantity of antigen.
If this seems too long, we also propose our Xpressway protocol of 28 days (Fast polyclonal antibody production) which is made using our special adjuvant which is aimed to increase the immune response.
It is usually only recommenced for anti-protein polyclonal antibodies because peptides might sometimes be not immunogenic enough to give a good immune response on this short period of time.
Species available at ProteoGenix
About animal species, rabbit polyclonal antibodies are usually the first choice for pab generation, however other species should be considered.
When looking for larger amounts of antiserum it may be more beneficial to use polyclonal antibodies from a larger animal such as sheep polyclonal antibody or goat polyclonal antibody.
Usually we advise to select an animal which is genetically distant from the antigen source (for example chicken polyclonal antibodies are very suitable to develop polyclonal antibodies against conserved mammalian proteins).
We also recommend our customers to use at least 2 animals per project. There is often a variability of immune response between individuals, so it increases the probability to get a highly reactive and specific polyclonal antibody targeting your antigen of interest.
Feel free to complete the online form above to design you own package !
Step 2: Select your
We offer up to seven different animal species for your polyclonal antibody preparation. A rabbit polyclonal antibody is the most common choice. Other species are useful for co-staining studies or for highly conserved target proteins, for example.
Step 3: choose your protocol
Select an animal species:
- GUINEA PIG
|Amount of serum / Final bleed (per animal)||Comment|
|Rabbit||30-40 ml of antiserum or 1 -10 mg of purified antibodies.||Rabbit polyclonal antibody is the first choice for antibodies generation.|
+ Best compromise in terms of cost and antibody quantity.
|Chicken||20-25 ml of antiserum/project or 1 -10 mg of purified antibodies.||Chicken antibody has higher phylogenetic distance to mammalian species.|
+ 5 to 10 times more antibodies than rabbits over the years, as antibodies are collected from egg yolk.
+ Chicken IgY antibodies usually generate less background than rabbit polyclonal antibodies when used in applications involving mammals.
|Guinea Pig||2.5-5 ml of antiserum or 1-10 mg of purified antibodies.||+ Immune response usually arises quickly and is strong.|
+ Low amount of antigen required.
- Very sensitive animal which must be handled with care.
- Low amounts of antibodies/serum.
|Rat||2.5-5 ml of antiserum or 1-10 mg of purified antibodies.||Often used for monoclonal antibodies as an alternative to mice.|
- Rat antibodies also provide a low amount of antibodies/serum.
|Mouse||300-500µl of antiserum or 10-100 µg of purified antibodies.||+ Good alternative when several antibody species are needed for multiple simultaneous staining.|
+ Strong immune responses
- Low amounts of antibodies/serum.
|Goat & Sheep||500-1000 ml of antiserum or >20mg of purified antibodies.||+ Goat antibody and sheep antibody gives large amounts of antiserum/antibodies.|
- More difficult to test pre-immune sera from several animals to choose the one showing the least background / cross reactivity with the tested samples.
- More expensive than rabbit polyclonal (for example).
How many animals would you like to inject ?
Depending on your deadlines, budget, and needs, we can adapt all the steps of the polyclonal antibody development process to perfectly meet your requirements. You can either provide us with a custom polyclonal antibody production protocol or follow our XpressWay protocol or standard with optional extension protocols.
- STANDARD - 51 DAYS
+ OPTIONAL EXTENSION
- XPRESSWAY - 28 DAYS
NOT FOR CHICKEN, GOAT AND SHEEP
Antibody final application:
- FLOW CYTOMETRY
- SANDWICH ELISA
All our polyclonal antibody production packages include guaranteed titers in ELISA and for anti protein polyclonal antibodies we can also guarantee positive WB results (see Package 3).
Step 4: select the antibody
- NO PURIFICATION
- ANTIGEN SPECIFICSingle
- IGG/Y TOTAL
- DEPLETION & PURIFICATION
SEQUENCE OR ANTIGEN INFORMATION (OPTIONAL):
ACCESSION NUMBER (OPTIONAL):
Antigen type: Protein
Antigen provided by: You
Protein localization: Cytoplasm
Chosen animal: Mouse
Number of animals for injections: 1
Protocol duration: Standard
Final antibody application: ELISA
Antibody purification: No purification