antigen / antibody immobilization
"Lewis, Ben FMC
Ben.Lewis at fmc.sa.gov.au
Wed Feb 19 17:57:40 EST 2003
Simple answers to your questions are as follows:
1) Can tag antibodies or recombinant proteins with tags (eg Myc/His-tags)
that have affinity for specific columns.
2) IF KEPT UNDER THE CORRECT CONDITIONS rxn's should not occur if the Ab is
lyophilised with a solid support.
3) Ab affinity can be as simple as eg. pyrimidine nucleic acids binding with
a triple bond. Usually this occurs in either a linear or quaternary
(topological) manner, depending on the type of Ab (ie. polyclonal Ab's
contain both epitope types).
However, the questions asked cannot be addressed in a manner by which you
would understand immediately. That is, you would need to do some additional
reading. Your best option is to get hold of a good biochemistry text (not
microbiology) such as 'Molecular Biology of the Cell', Alberts, Bray, Lewis,
Raff, Roberts and Watson, 'Principles of Biochemistry', Lehninger, Nelson
and Cox. There is also a good section on antigenic variation in 'Genes VII',
Hope this helps.
Benjamin Lewis BSc (Hons) GRACI
Department of Clinical Pharmacology,
Level 6, Flinders Medical Centre,
Flinders University School of Medicine,
South Australia 5042,
T +61 8 82044795
F +61 8 82045114
E ben.lewis at fmc.sa.gov.au
From: breddy at asu.edu [mailto:breddy at asu.edu]
Sent: Thursday, February 20, 2003 6:47 AM
To: methods at hgmp.mrc.ac.uk
Subject: antigen / antibody immobilization
My field of work is devices and we are working with molecule-device
hybrid structures.. i have little knowledge in microbiology.. i would
like to gain some understanding of protein binding to functionalized
1. best ways to bind antigen (or antibody ) or protein molecules to a
substrate like glass with SiOx surface, to substrates that are
parafinic in nature say surfaces attached with long chain alkanes etc.
(i haven't found good literature on it).. or suggest any other
substrate functionalization that would be ideal for such attachment ?
2. are antigen molecules and antibody molecules active even when
attached as above ? ie after drying in N2 atmosphere etc.. does an
antibody (suspended in solution) attack an antigen so attached
(antigen is attached to surface and dried) ? or does the antigen need
to be in solution ?
3. Selectivity of antibody attack, what is the mechanism of
identifying ? my concern is do they attack inorgonic or organic
substrates on which I attach antigen.. say substrates that are
parafinic, epoxy based, glass like etc. how are the charge transfers
during the reaction between antibody and antigen.. ?
comments ? experiences ? or references are welcome.. thanks in
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