Affibody-mediated transferrin depletion for proteomics applications.
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.
Grönwall
Caroline
C
Department of Molecular Biotechnology, School of Biotechnology, AlbaNova University Center, Royal Institute of Technology (KTH), Stockholm, Sweden.
Sjöberg
Anna
A
Ramström
Margareta
M
Höidén-Guthenberg
Ingmarie
I
Hober
Sophia
S
Jonasson
Per
P
Ståhl
Stefan
S
D015153:Blotting, Western; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000134:cerebrospinal fluid / Q000378:metabolism; D008024:Ligands; D019151:Peptide Library; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods; D040901:Proteomics / Q000379:methods*; D012709:Serum Albumin / Q000134:cerebrospinal fluid / Q000378:metabolism; D014168:Transferrin / Q000134:cerebrospinal fluid / Q000378:metabolism*
D007074:Immunoglobulin G; D008024:Ligands; D019151:Peptide Library; D012709:Serum Albumin; D014168:Transferrin