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Online monitoring of immunoaffinity-based depletion of high-abundance blood proteins by UV spectrophotometry using enhanced green fluorescence protein and FITC-labeled human serum albumin

Cited 7 time in Web of Science Cited 7 time in Scopus
Authors
Kim, Kyunggon; Yu, Jiyoung; Min, Hophil; Kim, Hyunsoo; Yu, Hyeong Gon; Kim, Youngsoo; Kim, Byungwook
Issue Date
2010-12-01
Publisher
BIOMED CENTRAL LTD
Citation
PROTEOME SCIENCE; Vol.8 ; -
Abstract
Background: The removal of high-abundance proteins from plasma is an efficient approach to investigating flow-through proteins for biomarker discovery studies. Most depletion methods are based on multiple immunoaffinity methods available commercially including LC columns and spin columns. Despite its usefulness, high-abundance depletion has an intrinsic problem, the sponge effect, which should be assessed during depletion experiments. Concurrently, the yield of depletion of high-abundance proteins must be monitored during the use of the depletion column. To date, there is no reasonable technique for measuring the recovery of flow-through proteins after depletion and assessing the capacity for capture of high-abundance proteins. Results: In this study, we developed a method of measuring recovery yields of a multiple affinity removal system column easily and rapidly using enhanced green fluorescence protein as an indicator of flow-through proteins. Also, we monitored the capture efficiency through depletion of a high-abundance protein, albumin labeled with fluorescein isothiocyanate. Conclusion: This simple method can be applied easily to common high-abundance protein depletion methods, effectively reducing experimental variations in biomarker discovery studies.
ISSN
1477-5956
Language
English
URI
http://hdl.handle.net/10371/77844
DOI
https://doi.org/10.1186/1477-5956-8-62
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College of Medicine/School of Medicine (의과대학/대학원)Biomedical Engineering (의공학전공)Journal Papers (저널논문_의공학전공)
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