From: Geoff=Howlett%BioChem%UNIMELB@muwaye.unimelb.EDU.AU
  To  : rasmb@bbri.eri.harvard.edu
  Date: Mon, 2 May 94 8:09:51 +1000

...no subject...

Hi

I enclose part of a reply from Preston Hensley re. standards.  The IL-8 
system sounds attractive.  I forward the message after having consulted with 
Preston and with a view to promoting discussion.  

Geoff H.

Message starts here:

In the mean time, I have what may be just what the doctor ordered.   The
molecule is IL-8.  It's mass is 8462 Da and it exhibits a reversible
monomer dimer equilibrium with Kd of 16 uM.  The system is slightly
temperature dependent.  We are in the process of determining the salt and
pH dependencies.  We have also characterized the association by both
experimental (titration) and theoretical calorimetry.  Everything fits.  The
molecule can be produced in 100 mg + quantities from E coli and the
purification requires only two columns.  We could supply small quantities for
a while, but if we want to make it generally available, we should contract out
the purification.  We have one MS ready to send out and are writing a more
complete version which should be out in the summer.

But before we jump the gun, I think we need to think things through a little.
We actually need several standards.
1- a pure monomer, ie lysozyme or some such.
2- a simple self-associating system (IL-8, for instance)
3- and A + B = C system (and mAb and myoglobin ... this could be followed at
two wavelengths and would also be good for BIAcore, ie put the mAb on the
surface.)
4- possible others.

We need systems that will accessible to as many technologies as possible, ie
Centrifuge
Calorimetry
BIAcore - IAsys
others?

Let's talk a little before we rush ahead on this.  What do you think?

- Preston             

end of message.