Naked DNA Immunizations

Brian Taylor jel-4 at bones.biochem.ualberta.ca
Thu May 19 20:44:57 EST 1994


Hi.  I've been working with Naked DNA Immunization rats in order to elicit
specific antisera to a toxic protein that has been incredibly difficult to
make using other prokaryotic and eukaryotic expression systems ( E.coli,
baculovirus, CHO cells, etc.)  I was hoping to circumvent the problem of
making lots of a toxic protein for inoculation by doing DNA immunizations
instead.  I've had some initial success with DNA immunizations using murine
IL-10 in rats, although this is a small secreted protein, whereas my
protein is large and anchored.

I'd like to do some control experiments to answer the following questions:

When doing DNA immunizations, which gives the strongest antibody response:

(a) surface anchored proteins
(b) secreted proteins
[(c) intracellular proteins]

Which is the best method of inoculation: ip, im, iv

Do adjuvants make a difference?

I was wondering if anyone could give me some suggestions as to what a good
control protein would be for these experiments.


(1) Gene encoding protein must be cloned (these are DNA immunizations)
(2) The protein itself should be readily available, and antibodies to it
(3) The protein should be non-toxic with no potential immunomodulatory
effects
    (unlike IL-10)
(4) It would be nice if it had naturally occuring secreted and anchored
forms,
    so I don't have to add signals and delete anchors.  This has caused
problems 
    in the past.

I've thought of a few and I am getting ready to do some experiments with
them,
but it would be nice to see if anyone else had some better ideas.

Thanks,

Brian Taylor
Dept of MMID
University of Alberta CANADA

email: jel-4 at bones.biochem.ualberta.ca



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