Peter G Mcpherson

US Patent:

54928397, Feb 20, 1996

Filed:

Jan 25, 1994

Appl. No.:

8/186435

Inventors:

Kevin P. Campbell - Iowa City IA
Derrick R. Witcher - Coralville IA
Peter McPherson - Branford CT
Steven D. Kahl - Fishers IN
John D. Windass - Wokingham, GB
Terence Lewis - Bracknell, GB
Philip Bentley - Bracknell, GB

Assignee:

University of Iowa Research Foundation - Iowa City IA

International Classification:

G01N 33567
G01N 33534
C07K 1476

US Classification:

436504

Abstract:

Disclosed is a novel photo-activatable derivative of ryanodine 9-hydroxy-21-(4-azidobenzoyloxy)-9-epiryanodine. This novel ryanodine derivative has been conjugated to keyhole limpet hemocyanin (KLH) and used for the production of antibodies with high affinity and specificity to ryanodine. A radioimmunoassay specific for ryanodine was developed using the anti-ryanodine antibodies, and a dissociation constant for ryanodine of 1 nM was determined. Also disclosed is the binding of a labeled, photoactivatable derivative of ryanodine (ABRy) with the ryanodine receptor of skeletal, cardiac and brain membranes. Digestion of the labeled ryanodine receptor revealed a labeled 76 kDa tryptic fragment which is critical for the formation of the high affinity ryanodine binding site.