Novel approach Alzheimer's disease biomarkers

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1 novel approach

1.1 theory
1.2 method
1.3 result





novel approach

recent studies focus on use of autoantibody, not biological markers future treatment. however, there various arguments whether autoantibody method provides reliable biomarker. number of reports show patients ad have lower levels of serum anti-ab antibodies healthy individuals, , others have argued level of anti-ab antibody may higher in ad. in order avoid provide solution discrepancy in existing data, dr. gustaw came novel method of dissociation sample.


theory

in biological fluids, antibodies , antigens in state of dynamic equilibrium between bound , unbound forms concentration-dependent. antigen masks antibody, obstructs accurate measurement of antibody-antigen detection. dr. gustow discovered novel way enhance antibody-antigen detection. using dissociation buffer (1.5% bovine serume albumin (bsa) , 0.2m glycine hcl ph2/5), dissociated antigen-antibody complexes. shown in figure below, in dissociated samples, unbound antigen-antibody complexes reveal increased disease state compared non-diseased state.


method

prepare dissociation buffer: 1.4% bovine serum albumin + 0.2m glycine-hcl, ph2.5
incubate ab42 20 minutes
dissolve ab42 in 500 ul dissociation buffer in microcon centrifugal device
incubate @ 23 °c (73 °f) 20 minutes
centrifuge 20 minutes @ 16,000 g @ 23 °c (73 °f)
invert filter , spin 3 minutes @ 2000 g
bring sample neutral ph 15-2ul 2.5m tris ph9
add elisa buffer (1.5% bsa , 0.05% tween 20 in phosphate buffered saline)
perform elisa analysis.

result

the white block represents non-dissociation data. black block represents dissociation data. elisa result shows, detection of antibody blocked addition of beta-amyloid when experiment performed without dissociation. following dissociation, level of antibody detected increased level control level of control.


he used same methodology in vivo examine sera collected ad patients. results, surprisingly, demonstrated significant increase in antibody titer. contradicts majority of studies arguing amyloid-beta antibody decreases in ad patients. non-dissociated sample follows widespread theory amyloid-beta decreases in ad patients. however, had proven non-dissociated sample fails bring out valid result. dissociated sample results show significant increases in ad patients, contradicts majority of previous studies.








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