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CALIFORNIA STATE SCIENCE FAIR


Use of Quartz Crystal Microbalance to Study Antibody Antigen Interactions


Objectives/Goals:

The Quartz Crystal Microbalance (QCMB) is a simple device. The adsorption of a
compound on the
quartz surface causes a decrease in the resonance frequency that is proportional
to the mass adsorbed. My
goal was to use the QCMB to prepare a biosensor capable of measuring baculovirus
particles.

Methods/Materials:

Self assembled monolayers (SAM's) were used to modify the surface of a quartz
crystal to enable the
attachment of proteins such as antibodies. Antibodies against the baculovirus
coat protein gp64 were
immobilized on a SAM that contained an N-hydroxysuccinimide(NHS) group.
Additionally the regeneration of the bound antibody was examined by screening a
series of reagents
which enabled me to identify a combination of these reagents that efficiently
disrupt the antibody antigen
interaction with out damaging the bound antibody. This enabled the repeated
reuse of the antibody.

Results:

Results show that the quartz crystal microbalance successfully and rapidly
detected the presence of the
baculovirus via the gp64 antigen. The biosensor shows a remarkably linear
response, in the range of 10E6
to 10E7 pfu per mL. On the positive side, this response is sensitive enough to
be useful in cell culture,
since the concentration of baculovirus produced in cell culture is typically in
the range of 10E7 to 10E9
pfu per mL. On the negative side, the unit is temperature-sensitive, often
requiring an hour or more to
stabilize before useful measurements can be obtained.
The cleaning study showed that a mixture of acids, salts, and EDTA could
completely regenerate the
antibody and permitted more that 10 uses without any apparent lose in antibody
capacity.

Conclusions/Discussion:

The target antigen chosen for this project, the baculovirus Autographa
californica, is of particular interest
to many in the pharmaceutical and pesticide industries. This virus is most
active against the alfalfa looper,
which destroys many crops. This project has demonstrated its applicability to
industry in that baculovirus
titers were obtained with the quartz crystal microbalance biosensor, in less
than a day, with ability to
re-use it numerous times. The capability of this biosensor could expedite
industry#s ability to produce the
virus or products made by them.

Summary Statement:

The preparation of a biosensor to baculoviruses using a quartz crystal
microbalance.