A: The progression of a catalytic immune response; B: molecular recognition of anions by silica bound sapphyrin
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Authors
Thomas, Richard Edward
Subjects
Advisors
Iverson, Brent L.
Sessler, Jonathan L.
Date of Issue
1994-08
Date
Publisher
Monterey, California. Naval Postgraduate School
Language
en_US
Abstract
Part A: This thesis describes the first detailed study of
how a catalytic immune response matures over a series of
immunizations. Specifically, three male New Zealand White rabbits
were immunized with a 4- (carboxy) benzyl phenyl phosphate hapten
conjugated to keyhole limpet hemocyanin (KLH) . The three rabbits
received an initial immunization with the immunogen, followed by
three immunizations at three week intervals. Serum samples were
obtained from each rabbit ten days following every immunization.
Pure polyclonal IgG antibodies were obtained by a multiple step isolation and purification process. The catalytic activity of each
sample was demonstrated by the rate enhancement and catalytic
efficiency of the hydrolysis of the corresponding carbonate
substrate. The results show a remarkable similarity in the immune
response maturation seen in two of the three rabbits. The third
rabbit was diseased and lagged slightly behind. Through the four
immunizations, the immune response continued to optimize due to an
apparent increase in the affinity of the polyclonal antibodies.This is quite significant, in that it provides the first
experimental evidence that affinity maturation is important for
antibody catalysis. We have thus begun to shed the first rays of
light inside the black box that produces catalytic antibodies. Part B: A novel sapphyrin-functionalized silica gel has
been used to study the anionic binding properties of the expanded
porphyrin sapphyrin. The preparation of the first generation of
this type of support is discussed. The silica-bound sapphyrin
displayed longer retention times under neutral, isochratic HPLC
conditions, for molecules with sterically accessible oxyanions. In
particular, the sapphyrin-functionalized silica gel showed a
remarkable preference for arsonates and phosphates. Furthermore,
mixtures of nucleotides and oligonucleotides were cleanly separated
according to the number of phosphate units they possessed. Thus,
not only does sapphyrin-bound silica gel provide an exquisite way to investigate anion binding specificities of sapphyrin, it also
provides a viable method to selectively separate and analyze anion
types. These types of modified silica gels may be useful for
solving present day environmental problems, as well as separating
and analyzing phosphorylated biologically relevant molecules.
Type
Thesis
Description
Series/Report No
Department
Chemistry
Organization
Identifiers
NPS Report Number
Sponsors
Funder
Format
125 leaves.
Citation
Distribution Statement
Rights
This publication is a work of the U.S. Government as defined in Title 17, United States Code, Section 101. Copyright protection is not available for this work in the United States.