PRF&L
Standard Protocols for Custom
Polyclonal Production
in Guinea
Pigs
STANDARD PROTOCOL for PROTEIN and PEPTIDE ANTIGENS
Printable
pdf format PRF&L Guinea Pig Standard Protocol for
Protein
and Peptide Antigens
| Day 0 |
Preimmune Bleed -
about 5ml plasma. Preimmune will be stored here at
-80 degrees C until the first immune bleed. Inject 50-100ug intramuscular using Complete
Freund's Adjuvant (CFA). We will inject intramuscular
and subcutaneous if the concentration of antigen is
less than 0.8mg/ml. We can also inject some of the
intradermal upon request. Acrylamide gel can only be
injected subcutaneous.
|
| Day 14 |
Inject
25-50ug intramuscular (or intradermal) in Incomplete Freund's Adjuvant (IFA) |
| Day 28 |
Inject
25-50ug intramuscular with IFA. |
| Day 42 |
Bleed - about 5ml plasma - The Tuesday following Day 42 we will ship via Federal Express the preimmune bleed and this first immune 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. 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
15-25ug intramuscular or subcutaneous
with IFA. We will usually do intramuscular
unless you specify subcutaneous. |
| Day 70 |
Bleed -We will do bleeds every 28 days unless you specify. The Tuesday following Day 70 we will ship via Federal Express this bleed. It is up to the investigator to test the bleed(s), 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 Guinea Pig Standard Protocol for
Fusion
Protein Antigens
| Day 0 |
Preimmune Bleed -
about 5ml plasma. Preimmune will be stored here at
-80 degrees C until the Day 63 bleed. Inject 50-100ug intramuscular using Complete
Freund's Adjuvant (CFA). We will inject intramuscular
and subcutaneous if the concentration of antigen is
less than 0.8mg/ml. We can also inject some of the
intradermal upon request. Acrylamide gel can only be
injected subcutaneous.
|
| Day 28 |
Inject
25-50ug intramuscular (or intradermal) in Incomplete Freund's Adjuvant (IFA) |
| Day 42 |
Inject
25-50ug intramuscular with IFA. |
| Day 56 |
Bleed - about 5ml plasma Inject
15ug intramuscular using IFA
Inject 15ug 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 |
Bleed. The Tuesday following Day 63 we will ship via Federal Express the Preimmune bleed the Day 56 bleed and this 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 intramuscular using IFA We will usually do
intramuscular unless you specify subcutaneous. |
| Day 84 |
Bleed. We will do these
bleeds every 28 days unless you specify. It is up to
the investigator to test the bleed, 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 15-25ug of antigen intramuscular or subcutaneous with IFA. We will do a 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. Production bleeds
with guinea pigs provide you with the same volume as any
other bleed as we only offer one size bleed with guinea
pigs. 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 plasma collected for
production bleeds depends on the weight and the general
health of the Guinea Pig. We generally take 10ml of blood
yielding 4 to 6ml of plasma. We have found that drawing
plasma gives a high usable yield for each bleed. However,
you may request that we bleed 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
|
