(728d) Serum Antibody Profiling of Celiac Disease Using Bacterial Display for In Vitro Diagnostics

Celiac disease
(CD) is an autoimmune disease of the small intestine affecting genetically
susceptible individuals following the ingestion of gluten, a protein found in
wheat. Although the combination of a serological test with a small intestinal
biopsy is a highly effective method for CD diagnosis, there are still critical
questions that need to be addressed with regards to early disease detection, identifying
additional trigger antigens, and discovering ways to distinguish patients that
will not improve with a gluten-free diet. A new serum antibody profiling
strategy has been optimized to selectively detect a pool of patients'
antibodies that test positive for two established serological tests for CD and
not detect antibodies from a control patient set. A group of 18 unique
mimotopes, or peptides that mimic antigenic determinants, were isolated after 5
rounds of enrichment by fluorescent activated cell sorting (FACS) of a
bacterial cell display library of random 15-mer peptides. In order to determine
which mimotopes had the highest predictive value for CD, the cross-reactivity
of these mimotopes was tested on an individual patient basis with the original
discovery patient cohort as well as a preliminary training set. A panel of 8
clones correctly classified 17 out of 20 patients as Celiac, while only
misclassifying 3 out of 20 controls. Additional CD patient cohorts will be
acquired from the clinics of leading CD investigators and our refined bacterial
display antibody profiling strategy will be applied to each set of serum
samples. After a sufficiently large group of candidate mimotopes are obtained
from repeated screenings, the bacterial cells will be printed on glass slides
in a novel microarray assay that has been developed in parallel. Bacterial cell
microarrays will provide a high-throughput method to characterize the
sensitivity and specificity of the CD detector during the training and
validation phases of the study and establish the detector is of clinical
relevance.