 |
 |
 |
 |
 |
In my extensive experience of making monoclonal antibodies to all kinds of
antigens including cell surface antigens, soluble proteins and haptens, it's
not easy to predict what is the best way to immunise, all the methods seem
to work most but not all of the time! However the really crucial step to get
right is the screening assay. I've written many times in the past the
philosophy that what you get is what you screen for but not necessarily what
you want! For example if you desire monoclonal antibodies which see a native
antigen on the cell surface and you have immunised with a fusion protein,
then just screening by ELISA on the immunogen is wrong and you obviously
want to do your screening on cells expressing the antigen. If you have the
antigen in different forms, eg as a fusion protein, on transfectants and
also as a native molecule, then use one to immunise and the others to screen
on. This can help to eliminate monoclonal antibodies against contaminants or
cross reacting antigens.
Mike Clark, mrc7@cam.ac.uk http://www.path.cam.ac.uk/~mrc7/
-- o/ \\ // || ,_ o Dr. M.R. Clark, Division of Immunology <\__,\\ // __o || / /\, Cambridge University, Dept. Pathology "> || _`\<,_ // \\ \> | Tennis Court Rd., Cambridge CB2 1QP ` || (_)/ (_) // \\ \_ Tel. 01223 333705 Fax. 01223 333875
|
|
|
|
|
|
CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web