Affibody molecule-mediated depletion of HSA and IgG using different buffer compositions: a 15 min protocol for parallel processing of 1-48 samples.
High-abundant plasma proteins pose a challenge in a large number of proteomics-based technologies. Depletion of these high-abundant proteins has proven to be a fruitful strategy to circumvent masking of lower-abundant proteins that could serve as valuable biomarker candidates. However, current strategies often do not meet the throughput requirements of large-scale proteomic studies. In the present paper, a flexible and parallelized method for the depletion of high-abundant proteins is described, allowing the removal of the two most abundant proteins from 48 blood-derived samples in less than 15 min using Affibody molecules as affinity ligands. A sample-processing platform like this should be suitable for a number of proteomics technologies; its flexibility in buffer composition allows for different types of downstream applications.
Eriksson
Cecilia
C
AlbaNova University Center, Department of Biotechnology, Royal Institute of Technology, SE-106 91 Stockholm, Sweden.
Schwenk
Jochen M
JM
Sjöberg
Anna
A
Hober
Sophia
S
D001774:Blood Chemical Analysis; D002021:Buffers; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007163:Immunosorbent Techniques; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods*
D002021:Buffers; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin