Discovery of Immune Reactive Human Proteins by High-Density Protein Arrays and Customized Validation of Potential Biomarkers by ELISA

2017 ◽  
pp. 11-21
Author(s):  
Tara K. Sigdel ◽  
Minnie. M. Sarwal
Keyword(s):  
High Density ◽  
Protein Arrays ◽  
Human Proteins ◽  
Potential Biomarkers

scholarly journals Identification of Novel Native Autoantigens in Rheumatoid Arthritis

Biomedicines ◽  
2020 ◽  
Vol 8 (6) ◽  
pp. 141 ◽  
Author(s):  
Thomas B. G. Poulsen ◽  
Dres Damgaard ◽  
Malene Møller Jørgensen ◽  
Ladislav Senolt ◽  
Jonathan M. Blackburn ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
High Density ◽  
Protein Array ◽  
Post Translational Modification ◽  
Native Proteins ◽  
Healthy Donors ◽  
Array Technology ◽  
Human Proteins ◽  
Patient Subgroups ◽  
Citrullinated Peptide

The majority of patients diagnosed with rheumatoid arthritis (RA) have developed autoantibodies against neoepitopes in proteins that have undergone post-translational modification, e.g., citrullination or carbamylation. There is growing evidence of their molecular relevance and their potential utility to improve diagnosis, patient stratification, and prognosis for precision medicine. Autoantibodies reacting to native proteins may also have a role in RA pathogenesis, however, their reactivity patterns remain much less studied. We hypothesized that a high-density protein array technology could shed light onto the normal and disease-related autoantibodies produced in healthy and RA patient subgroups. In an exploratory study, we investigated the global reactivity of autoantibodies in plasma pools from 15 anti-cyclic citrullinated peptide (CCP)-positive and 10 anti-CCP-negative RA patients and 10 healthy donors against more than 1600 native and unmodified human proteins using a high-density protein array. A total of 102 proteins recognized by IgG autoantibodies were identified, hereof 86 were recognized by antibodies from CCP-positive RA patients and 76 from anti-CCP-negative RA patients, but not by antibodies from healthy donors. Twenty-four of the identified autoantigens have previously been identified in synovial fluid. Multiple human proteins in their native conformation are recognized by autoantibodies from anti-CCP-positive as well as anti-CCP-negative RA patients.

scholarly journals Next-generation high-density self-assembling functional protein arrays

2008 ◽  
Vol 5 (6) ◽  
pp. 535-538 ◽  
Author(s):  
Niroshan Ramachandran ◽  
Jacob V Raphael ◽  
Eugenie Hainsworth ◽  
Gokhan Demirkan ◽  
Manuel G Fuentes ◽  
...  
Keyword(s):  
High Density ◽  
Next Generation ◽  
Functional Protein ◽  
Protein Arrays ◽  
Self Assembling

NANOSTRUCTURED DENDRIMER MODIFIED GLASS SURFACES FOR PROTEIN MICROARRAY

2007 ◽  
Vol 06 (02) ◽  
pp. 161-165 ◽  
Author(s):  
PARAYIL KUMARAN AJIKUMAR ◽  
JIN KIAT NG ◽  
JIM YANG LEE ◽  
GREGORY STEPHANOPOULOS ◽  
HENG-PHON TOO
Keyword(s):  
Glass Surface ◽  
Density Functional ◽  
Protein Microarray ◽  
Pamam Dendrimer ◽  
High Density ◽  
Protein Arrays ◽  
Nonspecific Adsorption ◽  
Cell Lysate ◽  
Modified Surface ◽  
Glass Surfaces

Carboxyl PAMAM dendrimer (3.5 generation) was covalently coupled onto amine modified glass surface to prepare a monolayer of high density functional linkers. Activation of the carboxyl PAMAM surface and the subsequent immobilization of antibodies resulted in a high density protein microarray as compared to linear carboxyl linker surface. In addition, fluorescent labeled cell lysate showed extremely low nonspecific adsorption to the PAMAM modified surface, comparable to inert PEG surface. Thus, the carboxyl PAMAM modified surface is ideal for the generation of high density protein arrays for the detection of dilute antigens with superior sensitivity.

Self-assembly of vesicle nanoarrays on Si: A potential route to high-density functional protein arrays

2007 ◽  
Vol 90 (3) ◽  
pp. 033901 ◽  
Author(s):  
C. S. Ramanujan ◽  
K. Sumitomo ◽  
M. R. R. de Planque ◽  
H. Hibino ◽  
K. Torimitsu ◽  
...  
Keyword(s):  
Self Assembly ◽  
Density Functional ◽  
High Density ◽  
Functional Protein ◽  
Protein Arrays

scholarly journals Referencing cross-reactivity of detection antibodies for protein array experiments

F1000Research ◽  
2016 ◽  
Vol 5 ◽  
pp. 73 ◽  
Author(s):  
Darragh Lemass ◽  
Richard O'Kennedy ◽  
Gregor S. Kijanka
Keyword(s):  
Specific Binding ◽  
Cross Reactivity ◽  
Protein Array ◽  
Igg Antibody ◽  
Western Blots ◽  
Protein Arrays ◽  
Rabbit Igg ◽  
Human Proteins ◽  
Antibody Repertoires ◽  
Secondary Antibodies

Protein arrays are frequently used to profile antibody repertoires in humans and animals. High-throughput protein array characterisation of complex antibody repertoires requires a platform-dependent, lot-to-lot validation of secondary detection antibodies. This article details the validation of an affinity-isolated anti-chicken IgY antibody produced in rabbit and a goat anti-rabbit IgG antibody conjugated with alkaline phosphatase using protein arrays consisting of 7,390 distinct human proteins. Probing protein arrays with secondary antibodies in absence of chicken serum revealed non-specific binding to 61 distinct human proteins. The cross-reactivity of the tested secondary detection antibodies points towards the necessity of platform-specific antibody characterisation studies for all secondary immunoreagents. Secondary antibody characterisation using protein arrays enables generation of reference lists of cross-reactive proteins, which can be then excluded from analysis in follow-up experiments. Furthermore, making such cross-reactivity lists accessible to the wider research community may help to interpret data generated by the same antibodies in applications not related to protein arrays such as immunoprecipitation, Western blots or other immunoassays.

High-Density Serum/Plasma Reverse Phase Protein Arrays

2017 ◽  
pp. 229-238 ◽  
Author(s):  
Cecilia Hellström ◽  
Tea Dodig-Crnković ◽  
Mun-Gwan Hong ◽  
Jochen M. Schwenk ◽  
Peter Nilsson ◽  
...  
Keyword(s):  
High Density ◽  
Reverse Phase ◽  
Protein Arrays ◽  
Phase Protein
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