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Injections must only be done with sterile needles and syringes. Freund’s should be used with g l a s s s y r i n g e s . T h e a m o u n t o f a n t i g e n t o b e i n j e c t e d w i l l v a r y , b u t i n g e n e r a l f o r r a b b i t s u s e

of 50-1000 ug is recommended. dose.

Booster doses are usually one half to equal the priming

The ideal adjuvant should be capable of helping to induce a high titer of high avidity antibodies while having few to no side effects. Unfortunately there is no single adjuvant

agent ideal

for

advantages

and

use with all antigens or in all situations.

disadvantages.

The

most

commonly

used

Each available

adjuvants

at

FSU

product are RIBI

has and

Freunds

complete/incomplete.

Please

review

the

document

‘Antibody

Production

  • -

    Adjuvants’

for

further

information.

In

addition

to

adjuvant

selection,

consider

whether

a

carrier

protein

should be used as a conjugate to enhance the immune response. While immunogenic by themselves, small molecular weight peptides (<15 KD) synthetic peptides may be poorly antigenic. These antigens may need

many antigens are and non-protein or to be coupled to a

carrier

protein

to

increase

their

immunogenicity.

Peptides

larger

than

25-30

KD

probably

do

not

need

conjugation.

Common

conjugating

proteins

include

keyhole

limpet

hemocyanin

(KLH), bovine immunogenicity

serum albumin, thyroglobulin and ovalbumin. In some cases, enhanced is achieved by covalently linking immunogens to liposomes prior to injection.

The well

choice of carrier should be based on its as its ability to be adequately coupled to

own ability to stimulate the desired antigen.

the

immune

response

as

For further information on preparation of antigens, refer to the UC Berkley ACUC Guidelines for Polyclonal Antibody Production in Laboratory Animals.

For scientific inappropriate.

reasons, the reuse of rabbits in other immunization protocols is usually For humane reasons, re-immunization with Freund's complete adjuvant is

inappropriate and only permitted with ACUC approval. However, because immunization and bleeding are non-invasive and low stress procedures, antibody producing rabbits may be used in other experiments not involving antibody production.

It is recommended that investigators collect a pre-immunization blood sample. On occasion, rabbits may have closely related antibodies to something in the rabbit’s environment or their feed which may be similar to the antigen under investigation.

Following completion of any and all procedures, the individual animal’s record must be updated. Observations to include: date, antigen and amount, adjuvant, route, sites, total volume, blood collection volume, sedation or anesthetics used, blood collection site. Observations of post procedural complications (not eating, localized swellings, ulcers, etc.) should be made as soon as noticed. Any treatment for such must be recorded in the animal’s record.

Standard Polyclonal Antibody Production Schedule for Rabbits: Note: Inappropriate handling of rabbits can result in life threatening injuries. Training on how to handle rabbits and to perform injection and blood collection procedures is available from LAR. Project personnel should demonstrate their ability to perform these procedures to the satisfaction of the attending veterinarian or a qualified individual designated by the attending veterinarian.

Wear appropriate PPE (lab coat, gloves and possibly protective ear wear) when handling rabbits to avoid induction of animal allergies.

Remove the rabbit from its cage and place it on a solid surface that is rough enough for the rabbit to

Revised April 2007

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