The 'reverse capture' autoantibody microarray:a native antigen-based platform for autoantibody profiling

Ehrlich, Joshua R.; Qin, Shuhao; Liu, Brian C-S

doi:10.1038/nprot.2006.66

Cited by 35 publications

(29 citation statements)

References 11 publications

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“…(30) presented a ‘reverse-capture microarray’ method based on a dual-antibody sandwich ELISA. Cancer cell lysates or tumor lysates are incubated with commercial antibody arrays so that each antigen is immobilized on a different spot in their native configurations.…”

Section: Profiling Of Tumor-associated Autoantibodiesmentioning

confidence: 99%

Tumor-associated autoantibodies as diagnostic and prognostic biomarkers

Heo

Bahk²,

Cho³

2012

BMB Reports

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In the process of tumorigenesis, normal cells are remodeled to cancer cells and protein expression patterns are changed to those of tumor cells. A newly formed tumor microenvironment elicits the immune system and, as a result, a humoral immune response takes place. Although the tumor antigens are undetectable in sera at the early stage of tumorigenesis, the nature of an antibody amplification response to antigens makes tumor-associated autoantibodies as promising early biomarkers in cancer diagnosis. Moreover, the recent development of proteomic techniques that make neo-epitopes of tumor-associated autoantigens discovered concomitantly has opened a new area of ‘immuno-proteomics’, which presents tumor-associated autoantibody signatures and confers information to redefine the process of tumorigenesis. In this article, the strategies recently used to identify and validate serum autoantibodies are outlined and tumor-associated antigens suggested until now as diagnostic/prognostic biomarkers in various tumor types are reviewed. Also, the meaning of autoantibody signatures and their clinical utility in personalized medicine are discussed. [BMB Reports 2012; 45(12): 677-685]

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Section: Profiling Of Tumor-associated Autoantibodiesmentioning

confidence: 99%

Tumor-associated autoantibodies as diagnostic and prognostic biomarkers

Heo

Bahk²,

Cho³

2012

BMB Reports

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“…In order to identify autoantibody signatures that might differentiate between prostate cancer and BPH, a customized version of our previously reported reverse capture protein microarray technology (6-8) was developed. From experiments using an initial set of over 500 cancer related antigens, as well as from literature searches and the Cancer Immunome Database (www2.licr.org/CancerImmunomeDB), a customized array containing 27 unique antigens was tested.…”

Section: Introductionmentioning

confidence: 99%

Autoantibody signatures as biomarkers to distinguish prostate cancer from benign prostatic hyperplasia in patients with increased serum prostate specific antigen

O'Rourke

DiJohnson

Caïazzo

et al. 2012

Clinica Chimica Acta

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Background Serum prostate specific antigen (PSA) concentrations lack the specificity to differentiate prostate cancer from benign prostate hyperplasia (BPH), resulting in unnecessary biopsies. We identified 5 autoantibody signatures to specific cancer targets which might be able to differentiate prostate cancer from BPH in patients with increased serum PSA. Methods To identify autoantibody signatures as biomarkers, a native antigen reverse capture microarray platform was used. Briefly, well-characterized monoclonal antibodies were arrayed onto nanoparticle slides to capture native antigens from prostate cancer cells. Prostate cancer patient serum samples (n=41) and BPH patient samples (collected starting at the time of initial diagnosis) with a mean follow-up of 6.56 y without the diagnosis of cancer (n=39) were obtained. One hundred micrograms of IgGs were purified and labeled with a Cy3 dye and incubated on the arrays. The arrays were scanned for fluorescence and the intensity was quantified. Receiver operating characteristic curves were produced and the area under the curve (AUC) was determined. Results Using our microarray platform, we identified autoantibody signatures capable of distinguishing between prostate cancer and BPH. The top 5 autoantibody signatures were TARDBP, TLN1, PARK7, LEDGF/PSIP1, and CALD1. Combining these signatures resulted in an AUC of 0.95 (sensitivity of 95% at 80% specificity) compared to AUC of 0.5 for serum concentration PSA (sensitivity of 12.2% at 80% specificity). Conclusion Our preliminary results showed that we were able to identify specific autoantibody signatures that can differentiate prostate cancer from BPH, and may result in the reduction of unnecessary biopsies in patients with increased serum PSA.

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“…14,15 By using human cells as a source of protein, the need for recombinant proteins is bypassed and, therefore, the detection of immunogenic epitopes that are not present in recombinant proteins is facilitated. 14,15 Therefore, this platform has the virtue of including post-translational modifications occurring in a cell type or tissue thought to be relevant to the disease, preserving and making available for antibody binding the tertiary and quaternary structures of proteins and complexes, multiplexing for simultaneous and rapid measurement of independent reactivities, and facilitating proteinprotein interaction mapping with native biological samples. This biomarker discovery platform was initially employed in "proof-of-principle" analyses to test the feasibility of autoantibody profiling.…”

Section: Reverse-capture Autoantibody Platform For Identification Of mentioning

confidence: 99%

“…14,15 This platform allows biologically relevant antigens to be immobilized in their native configuration. The "reverse capture" autoantibody microarray platform is based on the dual-antibody sandwich immunoassay of ELISA (Figure 2).…”

Section: Reverse-capture Autoantibody Platform For Identification Of mentioning

confidence: 99%

A Comprehensive Approach Toward Novel Serum Biomarkers for Benign Prostatic Hyperplasia: The MPSA Consortium

et al. 2008

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Purpose-Clinical benign prostatic hyperplasia (BPH) is primarily diagnosed based on a diverse array of progressive lower urinary tract symptoms (LUTS) and is likely distinct from histological BPH, which is detected by the presence of non-malignant proliferation of prostate cells but may or may not be associated with symptoms. Pharmacological management of LUTS has emerged as an effective initial treatment for clinical BPH due to the introduction of new drug therapies shown to be effective in recent large clinical trials. Despite advances in symptom management and research into BPH pathology, diagnostic strategies for prediction of BPH progression and response to drug modalities are lacking and questions remain as to the molecular differences underlying clinical (symptomatic) versus histological (non-symptomatic) BPH. Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. Results-To date the MPSA has identified a number of promising biomarkers and other molecular and cellular changes associated with BPH. NIH Public AccessConclusions-These findings and ongoing consortium discovery efforts have the potential to provide a greater understanding of the defects underlying disease pathology and may lead to the development of early and more effective pharmacological treatment strategies for BPH.

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The 'reverse capture' autoantibody microarray:a native antigen-based platform for autoantibody profiling

Cited by 35 publications

References 11 publications

Tumor-associated autoantibodies as diagnostic and prognostic biomarkers

Tumor-associated autoantibodies as diagnostic and prognostic biomarkers

Autoantibody signatures as biomarkers to distinguish prostate cancer from benign prostatic hyperplasia in patients with increased serum prostate specific antigen

A Comprehensive Approach Toward Novel Serum Biomarkers for Benign Prostatic Hyperplasia: The MPSA Consortium

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