Patent ID: 7528239
Filing Date: 2009-05-05
Classification: A61K,C07K

Abstract:
1. A composition comprising a plurality of isolated antibodies or antigen-binding fragments thereof, wherein each of said antibodies or antigen-binding fragments of said plurality specifically binds to an isolated and purified human TRAIL-R receptor protein wherein said isolated and purified human TRAIL-R receptor protein is a product made by the process comprising the steps of: (a) isolating plasma membranes from Jurkat cells; (b) solubilizing and homogenizing said isolated plasma membranes of step (a); (c) centrifuging said solubilized and homogenized isolated plasma membranes of step (b) to yield a plasma membrane extract and a pellet; (d) applying said plasma membrane extract of step (c) to an anti-octapeptide monoclonal antibody affinity chromatography column, whereby said column of step (d) adsorbs non-specifically bound material and wherein said octapeptide has the sequence presented in SEQ ID NO: 5; (e) applying column flow-through from step (d) to an octapeptide-TRAIL ligand affinity chromatography column, whereby said column of step (e) specifically binds said TRAIL-R receptor protein and wherein said octapeptide—TRAIL ligand is a fusion protein of said octapeptide having the sequence presented in SEQ ID NO:5 and TRAIL ligand; (f) eluting fractions with TRAIL ligand binding activity from said column of step (e); and, (g) purifying said fractions of step (f) by reverse-phase HPLC to yield said isolated and purified TRAIL-R receptor protein, wherein said isolated and purified TRAIL-R receptor protein has a molecular weight of about 50 to 55 kilodaltons as determined by SDS polyacrylamide gel electrophoresis, and comprises the amino acid sequence VPANEGD (amino acids 327-333 of SEQ ID NO: 2).