S-Space College of Medicine/School of Medicine (의과대학/대학원) Biomedical Engineering (의공학전공) Journal Papers (저널논문_의공학전공)
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
- Kim, Kyunggon; Yu, Jiyoung; Min, Hophil; Kim, Hyunsoo; Yu, Hyeong Gon; Kim, Youngsoo; Kim, Byungwook
- Issue Date
- BIOMED CENTRAL LTD
- PROTEOME SCIENCE; Vol.8 ; -
- 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.