Jan,
Are you dealing with a whole IgG or an Fab?
Most Fab-antigen complexes will not be affected by 5mM DTT.
Fab production makes use of reducing agents that allow the
antibody hinge region to become exposed to proteases.
If you need DTT to prevent your antigen from aggregating, try
to crystallize the Fab-antigen complex without worring about the
DTT.
Enrico.
On Thu, 22 Apr 2010 10:29:18 +0200, Jan Rash <jan...@googlemail.com> wrote:
Hi All,
I have a simple question about the complex formation between
macromolecular
complex and antibody. My protein is stable in the presence of the 5mM DTT
and under these conditions the reducing environment is too strong for the
antibody to survive. I am also now trying to check the stability of the
protein in lower molar concentration of DTT, but as DTT being a strong
reducing agent it might still pose a threat to the disintegrate the
antibody.
Does anybody have experience in handling protein-antibody complexes using
other reducing agents? Your answers and help in this regard will be
highly
appreciated.
Thanks,
Jan
--
Enrico A. Stura D.Phil. (Oxon) , Tel: 33 (0)1 69 08 4302 Office
Room 19, Bat.152, Tel: 33 (0)1 69 08 9449 Lab
LTMB, SIMOPRO, IBiTec-S, CE Saclay, 91191 Gif-sur-Yvette, FRANCE
http://www-dsv.cea.fr/en/institutes/institute-of-biology-and-technology-saclay-ibitec-s/unites-de-recherche/department-of-molecular-engineering-of-proteins-simopro/molecular-toxinology-and-biotechnology-laboratory-ltmb/crystallogenesis-e.-stura
http://www.chem.gla.ac.uk/protein/mirror/stura/index2.html
e-mail: est...@cea.fr Fax: 33 (0)1 69 08 90 71
