Ultradepletion of Human Plasma using Chicken Antibodies: A Proof of Concept Study

Human plasma arguably represents the most comprehensive version of the human proteome. Despite its immense theoretical discovery potential, plasma has many high and medium abundance proteins that mask low abundance protein disease biomarkers of relevance, making the discovery of novel diagnostic mar...

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Bibliographic Details
Published in:Journal of proteome research 2013-06, Vol.12 (6), p.2399-2413
Main Authors: Tan, Sock-Hwee, Mohamedali, Abidali, Kapur, Amit, Baker, Mark S
Format: Article
Language:English
Subjects:
Animals
Blood Proteins - administration & dosage
Blood Proteins - chemistry
Blood Proteins - immunology
Blood Proteins - isolation & purification
Chemical Fractionation - methods
Chickens
Chromatography, Affinity - methods
Electrophoresis, Gel, Two-Dimensional
Female
Humans
Immunization
Immunoglobulins - chemistry
Immunoglobulins - immunology
Immunoglobulins - isolation & purification
Immunoprecipitation - methods
Male
Protein Binding
Proteomics
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Summary:Human plasma arguably represents the most comprehensive version of the human proteome. Despite its immense theoretical discovery potential, plasma has many high and medium abundance proteins that mask low abundance protein disease biomarkers of relevance, making the discovery of novel diagnostic markers particularly difficult. Some form of protein depletion and/or fractionation is essential in order to detect markers of low abundance. Here, we describe a “proof of concept” two-pronged approach to immunodeplete abundant proteins from human plasma. The method, called API (Abundant Protein Immunodepletion), involves the fractionation of plasma using dual ion exchange columns (protein repetitive orthogonal offline fractionation (PROOF)) to simplify the proteome, the production of polyclonal IgY against each fraction and finally using the purified antibodies in a immunodepletion column. We explored the use of this product for immunodepletion of human plasma and identified a total of 165 nonredundant proteins after depletion. Of these, 38 proteins that were not previously identified in nondepleted plasma were now detected. It is envisaged that further optimization of the method as well as its cyclic implementation (by reinjecting depleted plasma into chickens for second round of antibody production) can make this technology highly robust, extremely cost-effective, and ideal for high throughput biomarker discovery.
ISSN:1535-3893
1535-3907
DOI:10.1021/pr3007182