RE: peptide oligo link
James Farmar (jfarmar@nybc.org)
Wed, 6 Oct 1999 09:54:05 -0400
Anne,
If you can spare an epsilon-amino group in your peptide (in forming an amide
bond) or add an amino group to your C-terminus, I recommend using
disuccinimidyl suberate (DSS) to link a 5'-amino or T-amino group of the
oligo to an amino group in your peptide. I had a lot of success linking
oligos to APase and the workup is easy and cheap. (Biotechniques 11, 588
(1991)). Remember, oligos have to be purified with TEAc (pH 7) buffers;
they don't survive 0.1% TFA.
There are options in the spacer portion of these homobifunctional
crosslinkers that can give more rigidity or distance.
AcCH2Br can work very well with some precautions (the cleavage cocktail
should not contain any thiols). We have used it to make multiple antigenic
peptides by the Robey and Fields and the Mayer approaches. Since your amino
terminus will have Fx, you will need to introduce a Lys(Mtt) or a similarly
differentiated amino group in order to add the AcCH2Br group on the resin.
Good Chemistry,
Jim Farmar
Laboratory of MicroChemistry, New York Blood Center