Download Anti-protein polyclonal antibodies

From Research to Discovery
Antibody Development Services
Anti-protein
polyclonal
antibodies
From STANDARD
to ALL INCLUSIVE
service
Protocols
adapted to your
requirements
Advice from
experts in
immunology
www.covalab.com
Approved
Health & Safety
procedures
STANDARD
IMMUNOGEN
One of the crucial steps during an immunisation project to produce a specific antibody is
the choice of the immunogen, which can be of various nature as shown below:
Soluble proteins (1)
Microorganisms
Others
In solution
Lyophilised
Within polyacrylamide gel
Inactivated virus
Inactivated bacteria
Inactivated yeast
Cell extracts
Tissue extracts
Others (please contact us)
Depending on the host animal species you choose to develop your
polyclonal antibody (see below), our immunisation protocols require
between 25 and 200 µg of immunogen (2) per injection and per animal.
Our optional ALL INCLUSIVE service (see last page) requires an
additional 100 µg of immunogen to perform ELISA monitoring and,
when required, antigen affinity purification.
(1) According to the size of your protein of interest, a coupling to a carrier protein may be required.
(2) This value may be adapted to your specific needs; in this case, we disclaim any responsibility for the ineffectiveness or toxic effects on the animals
HOST SPECIES
Rabbit
Mouse
Rat
Guinea pig
Goat
Chicken(3)
New Zealand
BALB/c
Wistar
Dunkin-Hartley
Alpine Chamoisée
Isa-brown
Immunogen
50-100 µg
Immunogen
50-100 µg
Immunogen
50-100 µg
Immunogen
50-100 µg
Immunogen
100-200 µg
Immunogen
25-50 µg
Test Bleeds: 5 to 15 mL
Final Bleed: 50 to 70 mL
Test Bleeds: 30 µL
Final Bleed: 0.5 mL
Test Bleeds: 100 µL
Final Bleed: 2.5 to 3 mL
Test Bleeds: 100 µL
Final Bleed: 5 to 6 mL
Test Bleeds: 6 to 12 mL
Final Bleed: 250 to 350 mL
Test Bleeds: 0.5 mL
Final Bleed: 6 to 8 mL
The most common host animal is the New Zealand white rabbit,
but we can propose various other species according to your specific
requirements.
We recommend to immunise at least two animals, due to the
variability of the immune response to a given antigen between
individuals.
(3) Chickens produce a unique class of immunoglobulins called IgY, which are functionally equivalent to mammalian IgG. Due to the phylogenic separation between
birds and mammals, chickens are more likely to produce an immune response against conserved proteins within the class of mammals. Moreover, as the structure of
the Fc region of chicken IgY is sufficiently different from mammalian IgG, the background signal in very sensitive immunological applications is dramatically reduced.
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D SERVICE
IMMUNISATION PROTOCOLS
On request, pre-immune sera can be tested in order to choose the
most suitable hosts. Indeed, test bleeds from 4 animals per project can
be tested in your specific conditions to select the best animal(s) to be
immunised.
Our standard protocols are optimised according to the host
species in order to ensure the good quality of the immune response.
Nevertheless, specific and personalised protocols can be carried out if
required.
Our expertise and active follow-up will guide you all along the protocol.
A provisional calendar, reminding
you the main steps of the protocol:
injections, bleeds and deliveries
53
67
D 4th injection
4th injection
81
88
D 90
D 3rd injection
D D 2nd injection
D D 1st injection
Final bleed and
eggs harvest
69
2nd test bleed
55
D 30
74
4th injection
1st test bleed
16
Final bleed
D 63
3rd injection
2nd test bleed
D 2nd injection
53
D 1st injection
D D 42
D 0
21
1st test bleed
All experiments are undertaken by experienced and authorised staff
following Health and Safety procedures, established according to the
French legislation governing the use of animals in experiments.
D 67
4th injection
Pre-immune
serum sample
Pre-immune
serum sample
Final bleed
D 56
D 3rd injection
39
2nd injection
2nd test bleed
D 28
D 14
D 1st injection
0
Final bleed
1st test bleed
D 0
Pre-immune
serum sample
42
3rd injection
2nd test bleed
D 2nd injection
39
1st injection
D D 28
D 14
1st test bleed
D 0
Pre-immune
serum sample
D 3rd injection
Final bleed
53
2nd injection
39
1st injection
D D 28
D 14
1st test bleed
D 0
Pre-immune
serum sample
The intermediate test bleeds at
the indicated dates, to run tests in
your specific conditions
D You receive:
New protocols are first submitted to an ethical committee for
agreement. Our animal house is registered under the reference
No C21 464 04 EA.
At the end of the immunisation protocol, you can:
Terminate the protocol and
request the final immune serum
OR
Extend the protocol and request:
additional injections
and / or
additional test bleeds
ALL INCLUSIVE SERVICE
Includes Standard and Additional Services
ANTI-PROTEIN ELISA
The sera immunoreactivity can be monitored by ELISA using a microtitre plate coated with the immunogenic protein. Our ALL INCLUSIVE service
includes ELISA for every sample we take during the immunisatin protocol.
Example of the evolution of a
serum immunoreactivity during
an immunisation protocol.
You receive:
The ELISA results, showing the serum immunoreactivity, as well as its evolution over the protocol. Alongside with the results of your own tests,
our expertise and our advice, you are able at each step to make the right decision whether to continue the project or not.
ANTIBODY PURIFICATION
Protein A/G or antigen affinity purification
For mammal polyclonal antibody development
projects, ALL INCLUSIVE service includes the
purification of IgG from up to 5 mL of the final immune
serum on a protein A or G column.
Upon request:
- larger volumes of serum (from 10 to more than
100 mL) can be purified
- antigen affinity purification of the serum can be
performed in order to dramatically increase its specificity.
Purification of IgY from egg yolk
For chicken polyclonal antibody development
projects, ALL INCLUSIVE service includes
the purification of IgY from 3 egg yolks per
chicken by selective precipitation followed by
ion exchange chromatography.
Upon request:
- we can store frozen separated egg yolks for
subsequent extraction,
- we can also purify up to 10 mL of the final serum.
You receive:
Your purified antibodies
in a glycine buffer(4).
A data report showing the conditions
and results of the purification.
(4) Glycine 0.1 M, Tris 1 M pH7.8, sodium azide 0.02%. If you do not wish azide to be added, please let us know on your order.
www.covalab.com
Covalab S.A.S. — 11 avenue Albert Einstein 69100 Villeurbanne — France
Phone: +33 (0) 437 654 230 — Fax: +33 (0) 437 289 416
E-mail: [email protected]