Chapter 8
Serum Proteomic Profiling in Rheumatoid Arthritis
by Antibody Suspension Bead Arrays
Lucı ´a Lourido, Rocı ´o Paz-Gonza ´ lez, Cristina Ruiz-Romero,
Peter Nilsson, and Francisco J. Blanco
Abstract
The versatility of protein microarrays provides researchers with a wide variety of possibilities to address
proteomic studies. Therefore, protein microarrays are becoming very useful tools to identify candidate
biomarkers in human body fluids for disease states such as rheumatoid arthritis (RA). In RA serum, there is
a high prevalence of rheumatoid factor (RF), which is an antibody with high specificity against Fc portion of
IgG. The presence of RF, in particular RF-IgM, has the great potential to interfere with antibody-based
immunoassays by nonspecifically binding capture antibodies. Because of this concern, we describe a
procedure to reduce the interference of RF-IgM on RA serum protein profiling approaches based on
multiplexed antibody suspension bead arrays.
Key words Antibody bead arrays, Serum, Rheumatoid arthritis, Interference, Biomarker discovery
1 Introduction
Bead-based protein array technology is an alternative method to
commonly used planar arrays, which enable high-throughput proteomic analysis of up to 384 proteins in 384 samples using only a
few microliters of sample. In antibody suspension bead arrays, the
antibodies are immobilized onto color-coded beads, and then these
are combined to create antibody microarrays for measuring
hundreds of samples per analysis. This technology has been
described to discover disease-associated protein profiles [1–
4]. However, the capture antibodies are vulnerable to bind endogenous antibodies as rheumatoid factor (RF), commonly present in
rheumatoid arthritis (RA) serum samples, which may interfere with
the assay measurements [5, 6]. Here, we describe a strategy to
reduce the damage caused by IgM RF in a serum proteomic
profiling assay based on antibody suspension bead array (Fig. 1).
Mo ´ nica Carrera and Jesu ´ s Mateos (eds.), Shotgun Proteomics: Methods and Protocols, Methods in Molecular Biology, vol. 2259,
https://doi.org/10.1007/978-1-0716-1178-4_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
143
Serum Proteomic Profiling in Rheumatoid Arthritis
by Antibody Suspension Bead Arrays
Lucı ´a Lourido, Rocı ´o Paz-Gonza ´ lez, Cristina Ruiz-Romero,
Peter Nilsson, and Francisco J. Blanco
Abstract
The versatility of protein microarrays provides researchers with a wide variety of possibilities to address
proteomic studies. Therefore, protein microarrays are becoming very useful tools to identify candidate
biomarkers in human body fluids for disease states such as rheumatoid arthritis (RA). In RA serum, there is
a high prevalence of rheumatoid factor (RF), which is an antibody with high specificity against Fc portion of
IgG. The presence of RF, in particular RF-IgM, has the great potential to interfere with antibody-based
immunoassays by nonspecifically binding capture antibodies. Because of this concern, we describe a
procedure to reduce the interference of RF-IgM on RA serum protein profiling approaches based on
multiplexed antibody suspension bead arrays.
Key words Antibody bead arrays, Serum, Rheumatoid arthritis, Interference, Biomarker discovery
1 Introduction
Bead-based protein array technology is an alternative method to
commonly used planar arrays, which enable high-throughput proteomic analysis of up to 384 proteins in 384 samples using only a
few microliters of sample. In antibody suspension bead arrays, the
antibodies are immobilized onto color-coded beads, and then these
are combined to create antibody microarrays for measuring
hundreds of samples per analysis. This technology has been
described to discover disease-associated protein profiles [1–
4]. However, the capture antibodies are vulnerable to bind endogenous antibodies as rheumatoid factor (RF), commonly present in
rheumatoid arthritis (RA) serum samples, which may interfere with
the assay measurements [5, 6]. Here, we describe a strategy to
reduce the damage caused by IgM RF in a serum proteomic
profiling assay based on antibody suspension bead array (Fig. 1).
Mo ´ nica Carrera and Jesu ´ s Mateos (eds.), Shotgun Proteomics: Methods and Protocols, Methods in Molecular Biology, vol. 2259,
https://doi.org/10.1007/978-1-0716-1178-4_8, © Springer Science+Business Media, LLC, part of Springer Nature 2021
143