US2011166330A1PendingUtilityA1

GPCR crystalization method using an antibody

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Assignee: KOBILKA BRIANPriority: Oct 15, 2007Filed: Feb 2, 2011Published: Jul 7, 2011
Est. expiryOct 15, 2027(~1.2 yrs left)· nominal 20-yr term from priority
Inventors:Brian KobilkaDaniel K. RohrerPeter BramsAsna Masood
C07K 2317/55C07K 14/723C07K 16/2869C07K 2317/34A61P 37/04C07K 2299/00C07K 2317/92
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Claims

Abstract

An antibody that specifically binds a three dimensional epitope on the IC3 loop of a GPCR is provided. The antibody may be employed in a method that comprises: contacting a GPCR with a monovalent version of the antibody binding conditions to form a complex; and crystallizing the complex.

Claims

exact text as granted — not AI-modified
1 . A monoclonal antibody that specifically binds a three dimensional epitope on the intracellular region (IC3) loop of a GPCR, wherein said monoclonal antibody is characterized in that it:
 a) stabilizes the relative conformations of the transmembrane region 5 (TM5) and transmembrane region 6 (TM6) of said GPCR,   b) facilitates crystallization of said GPCR.   
     
     
         2 . The monoclonal antibody of  claim 1 , wherein said antibody specifically binds the IC3 loop of the β2 adrenoreceptor. 
     
     
         3 . The monoclonal antibody of  claim 1 , wherein said antibody is a monovalent antibody. 
     
     
         4 - 17 . (canceled) 
     
     
         18 . The monoclonal antibody of  claim 1 , wherein said monoclonal antibody is a single chain antibody. 
     
     
         19 . The monoclonal antibody of  claim 1 , wherein said monoclonal antibody is a Fab fragment. 
     
     
         20 . The monoclonal antibody of  claim 1 , wherein said monoclonal antibody comprises a light chain variable domain of SEQ ID NO. 1 and a heavy chain variable domain of SEQ ID NO:2. 
     
     
         21 . The monoclonal antibody of  claim 1 , wherein said monoclonal antibody is made by
 a) reconstituting a GPCR in artificial phospholipid vesicles to make an antigen; and   b) immunizing an animal with said antigen.   
     
     
         22 . The monoclonal antibody of  claim 1 , wherein said monoclonal antibody is rabbit, mouse or chicken. 
     
     
         23 . The monoclonal antibody of  claim 1 , wherein said GPCR is a receptor for a biogenic amine. 
     
     
         24 . The monoclonal antibody of  claim 1 , wherein said GPCR is a dopamine receptor. 
     
     
         25 . The monoclonal antibody of  claim 1 , wherein said GPCR is not a β2-adrenergic receptor. 
     
     
         26 . The monoclonal antibody of  claim 1 , wherein said GPCR is a seratonin receptor. 
     
     
         27 . The monoclonal antibody of  claim 1 , wherein said GPCR is an adrenergic receptor. 
     
     
         28 . The monoclonal antibody of  claim 1 , wherein said GPCR is a melanocortin receptor subtype 4. 
     
     
         29 . The monoclonal antibody of  claim 1 , wherein said GPCR is a ghrelin receptor. 
     
     
         30 . The monoclonal antibody of  claim 1 , wherein said GPCR is a metabotropic glutamate receptor. 
     
     
         31 . The monoclonal antibody of  claim 1 , wherein said GPCR is a chemokine receptor.

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