Biology Reference
In-Depth Information
TABLE 1
Comparison of Different Technologies for Autoantibody Discovery
Sample
consumption
Antigen sources
Throughput
Multiplexity
Target identi
cation
cDNA expression
Phage plaque
Low. Rounds of plaque
screening.
Medium. Many plaques
can be screened on one
membrane lift.
Easy. DNA
sequencing.
High
Phage display
& microarray
Medium. Screening on
spotted phages.
High
Easy. DNA
sequencing.
High for panning.
Low for arrays.
Bacterial expression
& microarray
High. Bacterial colonies
are spotted on glass
slides.
High
Easy. DNA
sequencing.
Low
Cell lysates
2D Western
Low. 2D SDS-PAGE and
Western blot.
High. Hundreds of
spots on each gel.
Medium-hard. Mass
spec of positive spots.
Medium
Natural protein array
Medium. Thousands of
fractions obtained from
liquid phase separation
of cell lysates.
High
Medium-hard. Mass
spec of positive
fractions.
Low
Recombinant proteins
Puri
ed proteins
High
High
Easy
Low
In vitro expression
mixtures
High
High
Easy
Low
On-chip synthesized
proteins
High
High
Easy
Low
Synthetic peptides
Synthetic peptides/
peptoids on arrays
High
High
Easy for spatially
addressed arrays.
NMR analysis may
be needed
for peptoid arrays.
Low
Synthetic human
peptidome & phage
display
Low. Rounds of panning
and NextGen sequencing.
High
Hard. Sophisticated
analysis of NextGen
sequencing data.
Medium
Microbial surface
display
Low. Rounds of
panning.
High
Easy. DNA
sequencing.
Medium
systems are the ability to rapidly convert a target
library for screening low numbers of sera
for autoantigen discovery without
overrepresentation of abundant proteins, frame
shifts, and lower throughput have limited
the ultimate translation of these biomarkers into
clinical practice.
speci
c
equipment. However,
limitations such as the
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