CABELL.ANDREWS@BIONET-20.BIO.NET (Paul Andrews) (01/31/89)
Use of Sulfo-EGS as a Linker:
I am interested in using a homobifunctional NHS ester: Sulfo-EGS
[4-ethlyene glycoBIS-(sulfosuccininmidyl succinate)], to link a peptide (20 Aa
long) to albumin. Because the 20-mer by itself is too water soluble to be
precitated by TCA, linking the 20-mer to albumin should result in a complex
that is easily precipitated. I chose Sulfo-EGS (a new reagent from Pierce)
because it is water soluble, reacts on with primary amino groups, and is
easily cleaved with hydroxylamine. The whole purpose of this experiment is to
do some enzyme mediated coupling of a co-factor to a cysteine residue on the
20-mer. My problem is that I don't have a lot of this 20-mer (it is VERY
expensive) and don't want to waste a lot of it determining optimum conditions
for linking of the 20-mer to albumin. Sulfo-EGS is such a new reagent that
there are very few reports of its use in the literature.................
..............I was hoping that if someone has used this reagent as linker, or
has had experience in making peptide-protein links, they could send me a
protocol,.....or at least comment on whether this idea would work, or if
there are alternate methods of linking the 20-mer to a protein.
Thanks,
Paul Andrews
=============================================================================
Paul Andrews # EMAIL:CABELL.ANDREWS@BIONET-20.BIO.NET
# :CABELL.ANDREWS@[128.92.192.5]
University of Texas at Austin # Phone: (512) 471-3659, 471-1731
College of Pharmacy #
Division of Medicinal Chemistry #
Austin, Texas, USA
78712
-------wallace@fmi.ch (Andrew Wallace) (02/03/89)
Paul, If you are worried about wasting your peptide, why not use a tried- and-tested reagent like MBS which should work in the same way as Sulpho-EGS, i.e. linking amino groups on the carrier protein to a cysteine residue on the peptide. Is there some particular reason why you have chosen the new Pierce reagent? Andrew Wallace, FMI, Basel, Switzerland.