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Affibody‐mediated transferrin depletion for proteomics applications
Author(s) -
Grönwall Caroline,
Sjöberg Anna,
Ramström Margareta,
HöidénGuthenberg Ingmarie,
Hober Sophia,
Jonasson Per,
Ståhl Stefan
Publication year - 2007
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200700053
Subject(s) - transferrin , proteomics , chemistry , transferrin receptor , human plasma , blood proteins , western blot , biochemistry , microbiology and biotechnology , chromatography , biology , gene
An Affibody® (Affibody) ligand with specific binding to human transferrin was selected by phage display technology from a combinatorial protein library based on the staphylococcal protein A (SpA)‐derived Z domain. Strong and selective binding of the selected Affibody ligand to transferrin was demonstrated using biosensor technology and dot blot analysis. Impressive specificity was demonstrated as transferrin was the only protein recovered by affinity chromatography from human plasma. Efficient Affibody‐mediated capture of transferrin, combined with IgG‐ and HSA‐depletion, was demonstrated for human plasma and cerebrospinal fluid (CSF). For plasma, 85% of the total transferrin content in the samples was depleted after only two cycles of transferrin removal, and for CSF, 78% efficiency was obtained in single‐step depletion. These results clearly suggest a potential for the development of Affibody‐based resins for the removal of abundant proteins in proteomics analyses.