liposomes

Jim Owens jow at helix.nih.gov
Fri Jul 2 08:49:03 EST 1993


In article <9306282229.AA25896 at net.bio.net> Larry Anthony,
OLAF at UVVM.UVIC.CA writes:
>   I am trying to incorporate peptide antigens into
distearoylphosphatidylcholi
>ine/cholesterol unilamellar liposomes for immunization purposes. 
However, the
>protocol I have for making dehydration/rehydration vesicles (Kahl et al,
J. Imm
>unol. 142:4441, 1989) doesn't seem to give me any incorporation. 
Furthermore,
>the product apparently wont lyse in the presence of detergents,
including 5% 
>SDS.  Obviously I'm thinking that I'm doing something wrong, so I was
wondering 
>if anyone out there had a simple protocol for incorporating peptides into
>liposomes with this composition.

My wife is a whiz with liposomes.  She is Roberta Richards of the Walter
Reed Army Institute of Research.  She said not to be concerned if
detergents do not release your peptide antigens since they have probably
been incorporated into the bilayer.  This formulation of liposomes
usually resists dispersion of the bilayer in anything short of alcoholic
KOH.  She has taken your message to work where she can look up some good
references for your situation.  She recommended, in the meantime, that
you refer to an article:

Alving CR; Swartz GM Jr
Antibodies to cholesterol, cholesterol conjugates and liposomes:
implications for atherosclerosis and autoimmunity.
Department of Membrane Biochemistry, Walter Reed Army Institute of
Research, Washington, DC 20307-5100.
Language:  Eng
Source:  Crit Rev Immunol 1991;10(5):441-53

Another one that might be good is:
Alving CR
Liposomes as carriers of antigens and adjuvants.
Department of Membrane Biochemistry, Walter Reed Army Institute of
Research, Washington, DC 20307-5100.
Language:  Eng
Source:  J Immunol Methods 1991 Jun 24;140(1):1-13

Her group likes to use dimyristoylphosphatidylcholine instead of the
18-carbon chain fatty acid distearoylphosphatidylcholine.  I guess these
references will explain why.

Jim Owens



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