Comparison between monoclonal and polyclonal properties for custom antibodies synthesis
Monoclonal antibodies
Polyclonal antibodies
Specificity
A hybridoma will produce one single antibody type, detecting one single epitope present on the immunogen.Avoids cross-reactivity issues with other target proteins after a good clone is selected.
Possibility to select a clone that will be the most efficient in the given application, such as protein function inhibition, flow cytometry, sandwich ELISA …
Unpurified serum:Pool of antibodies, including non-antigen-specific antibodies, will bind different epitopes of the antigen.
Can generate background and cross-reactions with other proteins.
Affinity purified serum:Sample purity = 99.99%
Antibodies present in the serum are 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 antibodies can allow strongly binding ones to be found.
Sometimes more sensitive due to the wide variety of different antibodies represented in the serum.
Delay
It usually takes around 2 more months to get purified antibodies.
Usually a good alternative to get antibodies faster and cheaper for first tests.
Cost
3 to 4 times more expensive than the cost of polyclonal antibodies.
3 to 4 times less expensive than the cost of monoclonal antibodies.
Reproducibility
Antibodies always identicalMany lots of identical antibodies can be produced over dozens of years.
It is possible to sequence the hybridoma to make recombinant antibodies.
Antibodies different for each batch.Require repeated 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.
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As an experienced antibody manufacturer, ProteoGenix has successfully raised more than 1500 polyclonal antibodies during the past 13 years.
Step 1: Define your package
Antigen from customer 2 rabbits 1 purification
PACK 1
Anti-Peptide 2 rabbits 1 purification
PACK 2
Anti-Protein 2 rabbits 1 purification
PACK 3
Antigen from customer 2 rabbits No purification EXPRESS PROTOCOL
We offer up to seven different animal species for your antibody production. Rabbit is the preferred host species, while other species are useful for co-staining studies or for highly conserved target proteins.
>2.5 mg of protein or >2.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
30-40 ml of antiserum or 1 -10 mg of purified antibodies.
First choice for polyclonal.
Most preferred species for polyclonal antibody production. + Best compromise in terms of cost and antibody quantity.
Chicken
>2.5 mg of protein or >2.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
30-40 ml of antiserum/project or 1 -10 mg of purified antibodies.
Polyclonal against mammalian antigens.
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’s polyclonal antibodies when used in applications involving mammals.
Guinea Pig
>1.5 mg of protein or >1 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
2.5-5 ml of antiserum or 1-10 mg of purified antibodies.
Small volume. Polyclonal
+ Immune response usually arises quickly and is strong. + Low amount of antigen required. – Very sensitive animal which must be handled with care.
Rat
>1.5 mg of protein or >1 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
2.5-5 ml of antiserum or 1-10 mg of purified antibodies.
Small volume. Polyclonal or monoclonal.
Often used for monoclonal antibodies as an alternative to mice.
Mouse
>0.5 mg of protein or >0.5 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
300-500µl of antiserum or 10-100 µg of purified antibodies.
Small volume. Polyclonal or monoclonal.
+ Good alternative when several antibody species are needed for multiple simultaneous staining. – Strong immune responses, but low amounts of antibodies/serum.
Goat & Sheep
>10 mg of protein or >10 mg of conjugated peptide and 0.5mg of unconjugated peptide for ELISA testing.
500-1000 ml of antiserum or >100mg of purified antibodies.
Large volume. Polyclonal.
+ Large amounts of antiserum obtained. – 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.
* Protein (checked by SDS-PAGE), purity >90% Peptide for immunization has to be conjugated to a carrier (KLH, OVA or BSA) – we can also do the conjugation step if needed
How many animals would you like to inject ?
1
2
3
4
5
10
OTHER
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 antibody production protocol or follow our ExpressWay or standard with optional extension protocols.
Protocol duration:
STANDARD – 51 DAYS + OPTIONAL EXTENSION
EXPRESSWAY – 28 DAYS NOT FOR CHICKEN, GOAT AND SHEEP
CUSTOM
Antibody final application:
ELISA
WB
IHC
IF
FLOW CYTOMETRY
IP
SANDWICH ELISA
OTHER
All our polyclonal antibody production packages include guaranteed titers in ELISA > 1/64 000 guaranteed for purified antibodies.
Step 4: select the antibody purification method
How to choose the most appropriate type of purification ?
