Current applications of antibody microarrays
Chen et al. Clin Proteom
Current applications of antibody microarrays
Ziqing Chen 4
Tea Dodig‑Crnković 0 3
Jochen M. Schwenk 0 3
Sheng‑ce Tao 1 2 4 5
0 Affinity Proteomics, SciLifeLab, KTH ‐ Royal Institute of Technology , 171 65 Solna , Sweden
1 State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong University , Shanghai 200240 , China
2 School of Biomedical Engineering, Shanghai Jiao Tong University , Shanghai 200240 , China
3 Affinity Proteomics, SciLifeLab, KTH ‐ Royal Institute of Technology , 171 65 Solna , Sweden
4 Key Laboratory of Systems Biomedicine, (Ministry of Education), Shanghai Center for Systems Biomedicine, Shanghai Jiao Tong University , 800 Dong‐ chuan Road, Shanghai 200240 , China
5 State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiao Tong Univer‐ sity , Shanghai 200240 , China
The concept of antibody microarrays is one of the most versatile approaches within multiplexed immunoassay technologies. These types of arrays have increasingly become an attractive tool for the exploratory detection and study of protein abundance, function, pathways, and potential drug targets. Due to the properties of the antibody microarrays and their potential use in basic research and clinical analytics, various types of antibody microarrays have already been developed. In spite of the growing number of studies utilizing this technique, few reviews about antibody microarray technology have been presented to reflect the quality and future uses of the generated data. In this review, we provide a summary of the recent applications of antibody microarray techniques in basic biology and clinical studies, providing insights into the current trends and future of protein analysis.
Antibody microarray; Signalling; Drug mechanism; Clinical application; Systems biology; Technology advances
Background
Antibody microarrays are built on immobilizing
antibodies for a parallel analysis of multiple targets in a
given sample [
1
]. Today’s antibody and affinity
reagentengineering methods have helped to advance the
methodology [
2, 3
]. Antibodies and a variety of antibody
derivatives have been used to build arrays, including
nanobodies, single-chain variable fragments (scFvs) and
fragment antigen-binding (Fab)-fragments [4]. In addition,
phage display [
5
] and ribosome display [
6
], combined
with advanced materials and bioinformatics development
have being driving forces in recent years [
7
].
The typical workflow of an antibody microarray is
depicted in Fig. 1. Briefly, antibodies are immobilized
onto a chemically functionalized or otherwise modified
surface. After blocking the reactive groups of the
surface, a sample containing soluble proteins of interest is
incubated on the array, and the targeted proteins from
the sample are captured by the antibodies. The resulting
binding events are reported directly by fluorescent
labelling of the sample or by the addition of a secondary
detection reagent.
The attractiveness of antibody microarrays is that they
can be used to study a diverse number of biological
processes [
8
] and have been used to investigate
protein–protein interactions [
9
], signal pathway analysis [
10
], studies
of post-translation modifications [
11
], and detection of
toxins [
12
]. In the clinical context, arrays have enabled
opportunities to identify novel disease biomarkers [
13
] as
well as generating unique proteome signature by
comparing healthy and disease states. This information will be
of great value in the future, enabling better disease
management through improved diagnostics and the ability to
track disease status and therapeutic efficacy.
Antibody microarrays have demonstrated a number
of advantages compared to traditional, single analyte
methods of protein analysis, such as, enzyme-linked
immunosorbent assays (ELISA) and Western blotting.
Microarrays are high throughput, highly sensitivity,
require small sample volumes, and more recently have
become more standardized and user friendly
experimental procedures. Compared with mainstream proteomics
strategies, especially mass spectrometry (MS), the
process of antibody microarray assays is fast and takes less
than 24 h from sample preparation to data interpretation.
Detailed comparison is shown Fig. 2.
In theory, like DNA microarrays, antibody
microarrays can be designed to host a few to thousands, or even
ten-thousands, of features. Currently, high features have
been achieved by immobilizing proteins [
14
] or lysates
[
15, 16
], and antibody microarrays are under active
technological development and to-date operate at a few
hundred features [
17–19
]. The arrays can be constructed
either host many features per sample or be designed to
compartmentalize the array into sets of arrays that allow
many samples to be investigated simultaneously.
Generally, the latter is more common, particularly if a large
number of clinical samples are analysed in a given study.
For the analysis of (...truncated)