Ab's production from recombinant proteins - (Dec/03/2007 )

Hello friends!

I am working on recombinant proteins in order to produce matching Ab's. i would like suggestions or information

about production of Ab's from Denaturing proteins. can it be done?

yes, you can make antibodies with denatured protein.

are you preparing mono or polyclonal antibodies?

polyclonals will probably bind to native protein as well as denatured.

monoclonals, some clones will react with native and denatured, some only with denatured.

hi,
in agreement with medfenko, here is the explanation.

epitopes are of two types,

1-linear epitopes - Ab recongnizes short stretch of amino acids in a sequence (basically a primary structure).
2-confirmational epitopes - Ab recognize only when the protein is in native or partly denatured from (sec, ter or quatarnary structure).

u can obtain Abs against both 1 n 2 conditions.

mAb - as they are derived from single clone it can be categorized either into 1 or 2.
pAb - u have mixture of combinations (several B cell cones involved in the production) which recognizes both 1 and 2.

to answer ur question, if u denature ur protein u will exclude the 2nd possibility. hence at the end u will have Ab which recognizes only linear epitopes.


sravan payeli

Although polyclonal Abs binds denatured proteins in most cases. I've found that with recombinant proteins, polyclonal Abs sometimes fail to recognise antigen which was presented to the immune systen in another form ie native.

Thank u all for the information!