Design of recombinant antibody microarrays for urinary proteomics

Kristensson, Malin; Olsson, Karolina; Carlson, Joyce; Wullt, Björn; Sturfelt, Gunnar; Borrebaeck, Carl; Wingren, Christer

doi:10.1002/prca.201100055

Cited by 12 publications

(11 citation statements)

References 17 publications

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“…32 The specificity of several scFvs antibodies were previously validated using well-characterized, standardized serum samples (with known analytes of the targeted analytes), and orthogonal methods, such as mass spectrometry (affinity pull-down experiments), ELISA, MesoScale Discovery assay, cytometric bead assay, as well as using spike-in and blocking experiments. 19–21,33–38…”

Section: Methodsmentioning

confidence: 99%

Deciphering systemic lupus erythematosus-associated serum biomarkers reflecting apoptosis and disease activity

Delfani

Sturfelt

Gullstrand

et al. 2016

Lupus

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Systemic lupus erythematosus (SLE) is a severe chronic inflammatory autoimmune connective tissue disease. Despite major efforts, SLE remains a poorly understood disease with unpredictable course, unknown etiology and complex pathogenesis. Apoptosis combined with deficiency in clearing apoptotic cells is an important etiopathogenic event in SLE, which could contribute to the increased load of potential autoantigen(s); however, the lack of disease-specific protein signatures deciphering SLE and the underlying biological processes is striking and represents a key limitation. In this retrospective pilot study, we explored the immune system as a specific sensor for disease, in order to advance our understanding of SLE. To this end, we determined multiplexed serum protein expression profiles of crude SLE serum samples, using antibody microarrays. The aim was to identify differential immunoprofiles, or snapshots of the immune response modulated by the disease, reflecting apoptosis, a key process in the etiology of SLE and disease activity. The results showed that multiplexed panels of SLE-associated serum biomarkers could be decoded, in particular reflecting disease activity, but potentially the apoptosis process as well. While the former biomarkers could display a potential future use for prognosis, the latter biomarkers might help shed further light on the apoptosis process taking place in SLE.

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Section: Methodsmentioning

confidence: 99%

Deciphering systemic lupus erythematosus-associated serum biomarkers reflecting apoptosis and disease activity

Delfani

Sturfelt

Gullstrand

et al. 2016

Lupus

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“…Seventeen single-chain fragment variable antibodies were selected from in-house designed large phage-display libraries 22,23 . The specificity of antibodies from the libraries were previously validated with well-characterized serum samples (including spiking, blocking, and depletion of antigens) on antibody microarrays and several orthogonal methods such as mass spectrometry, enzyme-linked immunosorbent assay, and MesoScaleDiscovery cytokine assay, using various samples 16,18,[24][25][26][27][28][29][30] .…”

Section: Discussionmentioning

confidence: 99%

Multiplex profiling of serum proteins in solution using barcoded antibody fragments and next generation sequencing

et al. 2020

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The composition of serum proteins is reflecting the current health status and can, with the right tools, be used to detect early signs of disease, such as an emerging cancer. An earlier diagnosis of cancer would greatly increase the chance of an improved outcome for the patients. However, there is still an unmet need for proficient tools to decipher the information in the blood proteome, which calls for further technological development. Here, we present a proof-of-concept study that demonstrates an alternative approach for multiplexed protein profiling of serum samples in solution, using DNA barcoded scFv antibody fragments and next generation sequencing. The outcome shows high accuracy when discriminating samples derived from pancreatic cancer patients and healthy controls and represents a scalable alternative for serum analysis.

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“…The specificity, affinity, and onchip functionality have been assured using stringent phagedisplay screening and selection protocols using different sample formats ranging from pure proteins, mixtures of pure proteins to crude samples (Soderlind et al, 2000). In addition, the specificity of selected antibodies has been validated using pure proteins, mixtures of pure proteins, as well as wellcharacterized, standardized serum samples i) with known levels of the targeted analyte(s), ii) spiked with known level of specific protein(s), and/or iii) depleted of the targeted protein(s), and/or orthogonal methods such as mass spectrometry (affinity pull-down experiments), ELISA, Meso Scale Discovery assay and cytometric bead assay, as well as spiking and blocking experiments ( Supplementary Table 1) (Carlsson et al, 2011b;Dexlin-Mellby et al, 2010;Gustavsson et al, 2011;Ingvarsson et al, 2007Ingvarsson et al, , 2008Kristensson et al, 2012;Pauly et al, 2013;Wingren et al, 2007). Despite stringent selection and validation protocols, one limitation is the lack of information on fine specificity concerning proteoforms, translational modifications and potential complex formations.…”

Section: Generation Of Antibody Microarraysmentioning

confidence: 99%

Plasma protein profiling in a stage defined pancreatic cancer cohort – Implications for early diagnosis

et al. 2016

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Pancreatic ductal adenocarcinoma (PDAC) is a disease where detection preceding clinical symptoms significantly increases the life expectancy of patients. In this study, a recombinant antibody microarray platform was used to analyze 213 Chinese plasma samples from PDAC patients and normal control (NC) individuals. The cohort was stratified according to disease stage, i.e. resectable disease (stage I/II), locally advanced (stage III) and metastatic disease (stage IV). Support vector machine analysis showed that all PDAC stages could be discriminated from controls and that the accuracy increased with disease progression, from stage I to IV. Patients with stage I/II PDAC could be discriminated from NC with high accuracy based on a plasma protein signature, indicating a possibility for early diagnosis and increased detection rate of surgically resectable tumors.

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Design of recombinant antibody microarrays for urinary proteomics

Cited by 12 publications

References 17 publications

Deciphering systemic lupus erythematosus-associated serum biomarkers reflecting apoptosis and disease activity

Deciphering systemic lupus erythematosus-associated serum biomarkers reflecting apoptosis and disease activity

Multiplex profiling of serum proteins in solution using barcoded antibody fragments and next generation sequencing

Plasma protein profiling in a stage defined pancreatic cancer cohort – Implications for early diagnosis

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