Development of mammalian serum albumin affinity purification media by peptide phage display.
Several phage isolates that bind specifically to human serum albumin (HSA) were isolated from disulfide-constrained cyclic peptide phage-display libraries. The majority of corresponding synthetic peptides bind with micromolar affinity to HSA in low salt at pH 6.2, as determined by fluorescence anisotropy. One of the highest affinity peptides, DX-236, also bound well to several mammalian serum albumins (SA). Immobilized DX-236 quantitatively captures HSA from human serum; mild conditions (100 mM Tris, pH 9.1) allow release of HSA. The DX-236 affinity column bound HSA from human serum with a greater specificity than does Cibacron Blue agarose beads. In addition to its likely utility in HSA and other mammalian SA purifications, this peptide media may be useful in the proteomics and medical research markets for selective removal of mammalian albumin from serum prior to mass spectrometric and other analyses.
Sato
Aaron K
AK
Dyax Corp., One Kendall Square, Building 600, Suite 623, Cambridge, Massachusetts 02139, USA. asato@dynax.com
Sexton
Daniel J
DJ
Morganelli
Lee A
LA
Cohen
Edward H
EH
Wu
Qi Long
QL
Conley
Greg P
GP
Streltsova
Zoya
Z
Lee
Stan W
SW
Devlin
Mary
M
DeOliveira
Daniel B
DB
Enright
Jasmin
J
Kent
Rachel B
RB
Wescott
Charles R
CR
Ransohoff
Tom C
TC
Ley
Arthur C
AC
Ladner
Robert C
RC
D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000235:genetics / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D005454:Fluorescence Polarization / Q000379:methods; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D013045:Species Specificity
D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D012709:Serum Albumin