Performance evaluation of affinity ligands for depletion of abundant plasma proteins

Kullolli, Majlinda; Warren, Jonathan; Arampatzidou, Maria; Pitteri, Sharon J.

doi:10.1016/j.jchromb.2013.09.008

Cited by 26 publications

(20 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These kits are generally designated to be used for plasma samples and their application has been evaluated in several manuscripts (e.g. [ 22 , 24 , 26 – 28 ]). Kulloli et al [ 28 ] applied a kit for depletion of 14 abundant proteins in plasma prior to analysis by LC-MS/MS.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine

et al. 2015

View full text Add to dashboard Cite

Proteome analysis of complex biological samples for biomarker identification remains challenging, among others due to the extended range of protein concentrations. High-abundance proteins like albumin or IgG of plasma and urine, may interfere with the detection of potential disease biomarkers. Currently, several options are available for the depletion of abundant proteins in plasma. However, the applicability of these methods in urine has not been thoroughly investigated. In this study, we compared different, commercially available immunodepletion and ion-exchange based approaches on urine samples from both healthy subjects and CKD patients, for their reproducibility and efficiency in protein depletion. A starting urine volume of 500 μL was used to simulate conditions of a multi-institutional biomarker discovery study. All depletion approaches showed satisfactory reproducibility (n=5) in protein identification as well as protein abundance. Comparison of the depletion efficiency between the unfractionated and fractionated samples and the different depletion strategies, showed efficient depletion in all cases, with the exception of the ion-exchange kit. The depletion efficiency was found slightly higher in normal than in CKD samples and normal samples yielded more protein identifications than CKD samples when using both initial as well as corresponding depleted fractions. Along these lines, decrease in the amount of albumin and other targets as applicable, following depletion, was observed. Nevertheless, these depletion strategies did not yield a higher number of identifications in neither the urine from normal nor CKD patients. Collectively, when analyzing urine in the context of CKD biomarker identification, no added value of depletion strategies can be observed and analysis of unfractionated starting urine appears to be preferable.

show abstract

Section: Introductionmentioning

confidence: 99%

“…[ 22 , 24 , 26 – 28 ]). Kulloli et al [ 28 ] applied a kit for depletion of 14 abundant proteins in plasma prior to analysis by LC-MS/MS. The depletion allowed to enrich the sample for low-abundance proteins and increased the number of identifications compared to the non-depleted sample (from approx.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Removal of abundant plasma proteins by affinity antibodies is a common approach in this setting, as well (8). However, an alternative approach is to use mass spectrometry to study the low molecular weight peptides in plasma that represent intact protein degradation products.…”

Section: Proteomic Analysis Of Biological Fluids In the Study Of Kidnmentioning

confidence: 99%

Proteomics and autoimmune kidney disease

Rovin

Klein

2015

Clinical Immunology

View full text Add to dashboard Cite

Proteomics has long been considered an ideal platform, and urine an ideal source for biomarker discovery in human autoimmune kidney diseases. A number of studies have examined the urine proteome to identify biomarkers of disease activity, kidney pathology, and response to therapy. Increasingly, proteomic studies of kidney disease have expanded to include blood, circulating cells and kidney tissue. Recently the clinical potential of renal proteomics has been realized through a handful of investigations whose results appear to be applicable to patient care. In this review, approaches to the proteomic evaluation of autoimmune kidney diseases will be considered in the context of developing clinically useful disease biomarkers.

show abstract

“…Replacing traditional polyclonal antibodies with affinity ligands that do not require animal immunization and can be efficiently produced in large quantities lowers the cost of the depletion material. Examples of such ligands for FIB isolation are peptides or camelid antibody (Ab) fragments . To meet the demand of high‐throughput (HTP) glycomics studies, or glycan analysis of large numbers of samples, the methods used for analysis have to be fast, robust and reproducible.…”

Section: Introductionmentioning

confidence: 99%

Semi‐high‐throughput isolation and N‐glycan analysis of human fibrinogen using monolithic supports bearing monoclonal anti‐human fibrinogen antibodies

et al. 2017

View full text Add to dashboard Cite

Fibrinogen (FIB) is a secretory glycoprotein synthesized by hepatocytes that has a key role in blood clotting. Its glycosylation has not been studied in detail and little is known about the biological variability of FIB N-glycosylation, mainly due to the lack of fast, simple, and robust approaches to purify FIB from blood plasma samples. In recent years, customised chromatographic monoliths have been used for a variety of biological applications due to their unique characteristics. Here we describe development and optimisation of monolithic supports bearing monoclonal anti-human fibrinogen antibodies in a single column as well as in multi-well plate formats with high FIB specificity and binding capacity for fast immunoaffinity purification of FIB from human blood samples. The developed semi-high-throughput workflow has been successfully applied for FIB immunoaffinity isolation and subsequent ultra performance liquid chromatography N-glycosylation analysis in ten healthy human individuals, demonstrating the potential of monolithic supports in glycomics studies.

show abstract

Performance evaluation of affinity ligands for depletion of abundant plasma proteins

Cited by 26 publications

References 29 publications

Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine

Comparison of Depletion Strategies for the Enrichment of Low-Abundance Proteins in Urine

Proteomics and autoimmune kidney disease

Semi‐high‐throughput isolation and N‐glycan analysis of human fibrinogen using monolithic supports bearing monoclonal anti‐human fibrinogen antibodies

Contact Info

Product

Resources

About