|
PRF&L Standard Protocols for Custom
Polyclonal Production
in
Goats and Sheep
STANDARD PROTOCOL for PROTEIN and PEPTIDE ANTIGENS
Printable
pdf format PRF&L Goat Standard Protocol for
Protein
and Peptide Antigens
Printable
pdf format PRF&L Sheep Standard Protocol for
Protein
and Peptide Antigens
| Day 0 |
Test (5ml sera),
Small (60ml sera), or Large (200ml sera) Preimmune Bleed. Preimmune will be stored here at -80 degrees C
until the first immune bleed. Inject
100-200ug intradermal or intramuscular, or both, in Complete Freund's Adjuvant (CFA). We can also inject
into the popliteal lymph node upon request. We will inject intradermal and intramuscular if the
concentration of antigen is less than 1mg/ml. Acrylamide gel can only be injected subcutaneous.
|
| Day 14 |
Inject 50-100ug intradermal
or intramuscular
in Incomplete Freund's Adjuvant (IFA)
|
| Day 28 |
Inject 50-100ug
intramuscular
or intradermal (if we injected lymph node on Day 0) with IFA.
|
| Day 42 |
Small Bleed (60ml sera)
The Tuesday following Day 42 we will ship via Federal Express the preimmune bleed and this first small
bleed on dry ice. It is then up to the investigator to test the serum, determine the quality of antibodies,
and let us know if any changes are necessary in the protocol. If you prefer, we can take the test
bleed size (5ml).
PRF&L now offers
antibody titration by ELISA
and can now check titers for you.
There is an additional charge for this service.
|
| Day 56 |
Inject 25-50ug
intramuscular
with IFA
|
| Day 70 |
Small Bleed (3-5ml sera) We will
do small bleeds every 28 days unless you specify. The Tuesday following Day 70 we will ship via Federal
Express this small bleed. It is up to the investigator to test the serum, determine the quality of antibodies,
and let us know if any changes are necessary in the protocol.
PRF&L now offers
antibody titration by ELISA
and can now check titers for you.
There is an additional charge for this service.
|
| After Day 70 |
See Continuing your Protocol below. |
STANDARD PROTOCOL for FUSION PROTEIN ANTIGENS
Printable
pdf format PRF&L Goat Standard Protocol for
Fusion Protein
Antigens
Printable
pdf format PRF&L Sheep Standard Protocol for
Fusion Protein
Antigens
| Day 0 |
Test (5ml sera),
Small (60ml sera), or Large (200ml sera) Preimmune Bleed. Preimmune will be stored here at -80 degrees C
until the first immune bleed. Inject
100-200ug intradermal or intramuscular, or both, in Complete Freund's Adjuvant (CFA). We can also inject
into the popliteal lymph node upon request. We will inject intradermal and intramuscular if the
concentration of antigen is less than 1mg/ml. Acrylamide gel can only be injected subcutaneous.
|
| Day 28 |
Inject 50-100ug intradermal
or intramuscular
in Incomplete Freund's Adjuvant (IFA)
|
| Day 42 |
Inject 50-100ug subcutaneous or intradermal (if
we injected lymph node on day 0) with IFA.
|
| Day 56 |
Test Bleed (5ml sera)
Inject 25ug
or intramuscular
using IFA
Inject 25ug intravenous (antigen must be soluble
in PBS or water) using no adjuvant. If this is not available, we will skip the IV injection.
|
| Day 63 |
Large Bleed (200 to 225ml sera).
The Tuesday following Day 63 we will ship via Federal Express the Preimmune bleed the Day 56 bleed and
this large bleed. It is then up to the investigator to test the serum, determine the quality of antibodies,
and let us know if any changes are necessary in the protocol.
PRF&L now offers antibody titration by ELISA and can now check titers for you. There is
an additional charge for this service.
|
| Day 70 |
Inject 25-50ug
or intramuscular
using IFA
|
| Day 84 |
Small Bleed (60ml) We will
do these bleeds every 28 days unless you specify. It is up to the investigator to test the serum,
determine the quality of antibodies, and let us know if any changes are necessary in the protocol.
PRF&L now offers antibody
titration by ELISA and can now check titers for you. There is an additional charge for this service.
|
| After Day 84 |
See Continuing your Protocol below. |
CONTINUING for BOTH PROTOCOLS
| We will continue give boosts every
four weeks with 25-50ug of antigen intramuscular with IFA. We will do a small bleed two weeks after
each boost and send to you the following Tuesday. We will continue to boost and bleed every four
weeks until you contact us. Keep in mind that you have several other options depending on your
needs and the success of the project.
|
| When the antisera contains sufficient
antibody titer, we can start production bleeds. We can do production bleeds every seven days. Please contact us if
you want to change the bleeding and/or boosting schedule. You may also request that we not do any boosts after the
initial immunization scheme outlined above since some antigens and some animals may respond negatively to
boosts during the period of peak antibody activity. Boosts may be required only when titer drops. It is up to
the investigator to decide and let us know if we should discontinue boosts as our standard schedule provides for
boosts every 28 days.
|
| The volume of serum collected for
large or production bleeds depends on the weight and the general health of the Goat or Sheep. We generally take
400ml to 800ml of blood yielding 200 to 300ml of serum.
|
| Our experience shows that the peak
antibody titer occurs at an average of 120 days, however we have experienced the onset of peak activity as early
as six weeks and as long as a year. If results are not as you expect, please consult with us and we can make
recommendations on how to change your protocol.
|
| When you have collected enough
antisera, you will need to contact us to terminate the project. You may request that we perform an
exsanguination (terminal bleed) or if no additional sera is required, the animal(s) will be humanely euthanized
(discard). When you instruct us to terminate your animal(s), you must tell us if you want exsanguination or
discard, species, and the animal(s) number. Written confirmation by email or fax may be requested for
terminal procedures.
|
Select another Species
|
