PMID- 19683003
OWN - NLM
STAT- MEDLINE
DCOM- 20091027
LR  - 20091012
IS  - 1089-8638 (Electronic)
IS  - 0022-2836 (Linking)
VI  - 393
IP  - 3
DP  - 2009 Oct 30
TI  - DARPins as bispecific receptor antagonists analyzed for immunoglobulin E receptor 
      blockage.
PG  - 598-607
LID - 10.1016/j.jmb.2009.08.014 [doi]
AB  - The concept of multispecific antibodies is of high therapeutic interest but has 
      failed to produce pharmaceutical products due to the poor biophysical properties 
      of such molecules. Here, we propose an alternative and simple way to generate 
      bispecific binding molecules using designed ankyrin repeat proteins (DARPins). 
      For this purpose, monovalent DARPins with different epitope specificities were 
      selected against the alpha chain of the high-affinity receptor for human 
      immunoglobulin E (IgE) (FcepsilonRIalpha). Two of the isolated binders 
      interfering with IgE binding to the receptor were joined to each other or to 
      themselves via a flexible protein linker. The resulting bivalent and bispecific 
      DARPins were tested for their ability to prevent allergen-induced cell 
      degranulation using rat basophilic leukemia cells stably transfected with human 
      FcepsilonRIalpha. The bispecific DARPin construct was the most potent one, 
      efficiently blocking the IgE-FcepsilonRI interaction and preventing the release 
      of proinflammatory mediators. Noteworthy, the multivalent and multispecific 
      DARPin construct did not show any alteration of the beneficial biophysical 
      properties of the monovalent parental DARPins. Hence, bispecific DARPins may be 
      used to generate receptor antagonists simultaneously targeting different epitopes 
      on the same molecule. Moreover, they easily overcome the limiting immunoglobulin 
      binding paradigm (one binding molecule=one epitope) and thereby represent an 
      alternative to monoclonal antibodies in cases where the immunoglobulin scaffold 
      is unsuitable.
FAU - Eggel, Alexander
AU  - Eggel A
AD  - Institute of Immunology, University of Bern, Inselspital, Sahlihaus 2, Bern, 
      Switzerland.
FAU - Baumann, Michael J
AU  - Baumann MJ
FAU - Amstutz, Patrick
AU  - Amstutz P
FAU - Stadler, Beda M
AU  - Stadler BM
FAU - Vogel, Monique
AU  - Vogel M
LA  - eng
PT  - Journal Article
DEP - 20090813
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Epitopes)
RN  - 0 (FcepsilonRI alpha-chain, human)
RN  - 0 (Proteins)
RN  - 0 (Receptors, IgE)
SB  - IM
MH  - Anaphylaxis/immunology
MH  - Animals
MH  - *Ankyrin Repeat
MH  - Antibody Affinity
MH  - Antibody Specificity
MH  - Cell Line, Tumor
MH  - Chromatography, High Pressure Liquid
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Epitopes/immunology
MH  - Humans
MH  - Protein Structure, Quaternary
MH  - Proteins/immunology/*pharmacology
MH  - Rats
MH  - Receptors, IgE/*antagonists & inhibitors/immunology
MH  - Transfection
EDAT- 2009/08/18 09:00
MHDA- 2009/10/29 06:00
CRDT- 2009/08/18 09:00
PHST- 2009/06/18 00:00 [received]
PHST- 2009/08/06 00:00 [revised]
PHST- 2009/08/07 00:00 [accepted]
PHST- 2009/08/18 09:00 [entrez]
PHST- 2009/08/18 09:00 [pubmed]
PHST- 2009/10/29 06:00 [medline]
AID - S0022-2836(09)00998-X [pii]
AID - 10.1016/j.jmb.2009.08.014 [doi]
PST - ppublish
SO  - J Mol Biol. 2009 Oct 30;393(3):598-607. doi: 10.1016/j.jmb.2009.08.014. Epub 2009 
      Aug 13.