PMID- 9255793
OWN - NLM
STAT- MEDLINE
DCOM- 19971014
LR  - 20111117
IS  - 1087-0156 (Print)
IS  - 1087-0156 (Linking)
VI  - 15
IP  - 8
DP  - 1997 Aug
TI  - Binding proteins selected from combinatorial libraries of an alpha-helical 
      bacterial receptor domain.
PG  - 772-7
AB  - Small protein domains, capable of specific binding to different target proteins 
      have been selected using combinatorial approaches. These binding proteins, called 
      affibodies, were designed by randomization of 13 solvent-accessible surface 
      residues of a stable alpha-helical bacterial receptor domain Z, derived from 
      staphylococcal protein A. Repertoires of mutant Z domain genes were assembled and 
      inserted into a phagemid vector adapted for monovalent phage display. Two 
      libraries, each comprising approximately 4 x 10(7) transformants, were 
      constructed using either an NN(G/T) or an alternative (C/A/G)NN degeneracy. 
      Biopanning against the target proteins Taq DNA polymerase, human insulin, and a 
      human apolipoprotein A-1 variant, showed that in all cases significant 
      enrichments were obtained by the selection procedures. Selected clones were 
      subsequently expressed in Escherichia coli and analyzed by SDS-PAGE, circular 
      dichroism spectroscopy, and binding studies to their respective targets by 
      biospecific interaction analysis. The affibodies have a secondary structure 
      similar to the native Z domain and have micromolar dissociation constants (KD) 
      for their respective targets.
FAU - Nord, K
AU  - Nord K
AD  - Department of Biochemistry and Biotechnology, Royal Institute of Technology 
      (KTH), Stockholm, Sweden.
FAU - Gunneriusson, E
AU  - Gunneriusson E
FAU - Ringdahl, J
AU  - Ringdahl J
FAU - Stahl, S
AU  - Stahl S
FAU - Uhlen, M
AU  - Uhlen M
FAU - Nygren, P A
AU  - Nygren PA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Nat Biotechnol
JT  - Nature biotechnology
JID - 9604648
RN  - 0 (Apolipoprotein A-I)
RN  - 0 (Immunoglobulin Fc Fragments)
RN  - 0 (Insulin)
RN  - 0 (Peptide Library)
RN  - 0 (Receptors, Immunologic)
RN  - 0 (Staph protein A receptor)
RN  - 0 (Staphylococcal Protein A)
RN  - EC 2.7.7.- (Taq Polymerase)
RN  - EC 2.7.7.7 (DNA-Directed DNA Polymerase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Apolipoprotein A-I/metabolism
MH  - Binding Sites/genetics
MH  - Biosensing Techniques
MH  - Circular Dichroism
MH  - DNA-Directed DNA Polymerase/metabolism
MH  - Drug Design
MH  - Escherichia coli
MH  - Humans
MH  - Immunoglobulin Fc Fragments/*metabolism
MH  - Insulin/metabolism
MH  - Magnetic Resonance Spectroscopy
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - *Peptide Library
MH  - Protein Binding
MH  - Protein Conformation
MH  - Protein Structure, Secondary
MH  - Receptors, Immunologic/chemistry/genetics/*metabolism
MH  - Sequence Alignment
MH  - Staphylococcal Protein A/chemistry/genetics/*metabolism
MH  - Taq Polymerase
EDAT- 1997/08/01 00:00
MHDA- 1997/08/01 00:01
CRDT- 1997/08/01 00:00
PHST- 1997/08/01 00:00 [pubmed]
PHST- 1997/08/01 00:01 [medline]
PHST- 1997/08/01 00:00 [entrez]
AID - 10.1038/nbt0897-772 [doi]
PST - ppublish
SO  - Nat Biotechnol. 1997 Aug;15(8):772-7. doi: 10.1038/nbt0897-772.