       Document 0321
 DOCN  M9640321
 TI    A general method to recondition and reuse BIAcore sensor chips fouled
       with covalently immobilized protein/peptide.
 DT    9604
 AU    Chatelier RC; Gengenbach TR; Griesser HJ; Brigham-Burke M; O'Shannessy
       DJ; Division of Chemicals and Polymers, CSIRO, Clayton, Victoria,;
       Australia.
 SO    Anal Biochem. 1995 Jul 20;229(1):112-8. Unique Identifier : AIDSLINE
       MED/96140938
 AB    Of significance in the routine use of BIAcore is the cost of the sensor
       chips. This is particularly evident during the phase of method
       development of an assay where it is not unusual to expend several chips
       in a day in attempts to optimize immobilization conditions for a novel
       peptide or protein. In addition, it is accepted practice to discard a
       chip once its ligand binding capacity has diminished to an unacceptable
       level. While the high cost of sensor chips has been addressed to some
       degree through the recent introduction of research-grade sensor chips,
       we were interested in assessing the possibility of regenerating or
       reconditioning sensor chips in order to allow them to be reused. In
       particular, we concerned ourselves with regenerating sensor chips onto
       which peptide or protein had been immobilized. Our aim was to develop a
       general procedure that would allow reuse of such chips but would not
       decrease ligand immobilization capacity or increase nonspecific ligand
       adsorption properties. We present a method which employs a combination
       of enzymatic (Pronase E) and chemical (bromoacetic acid) treatments of
       used sensor chips. Regeneration requires an overnight incubation of the
       sensor chip ex situ so that one can continue to perform BIAcore
       experiments. The data demonstrate that this simple two-step procedure
       substantially removes immobilized proteins such as IgG, Protein G, an
       HIV-1 envelope glycoprotein (gp 120) and a neoglycoprotein based on
       bovine serum albumin, as determined by reflectance measurements and
       X-ray photoelectron spectroscopy.(ABSTRACT TRUNCATED AT 250 WORDS)
 DE    Animal  *Biosensors  Biotechnology  Cattle  Ligands  Methods
       Peptides/ISOLATION & PURIF  Pronase  Proteins/ISOLATION & PURIF
       Spectrometry, X-Ray Emission  Surface Properties  Time Factors  JOURNAL
       ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

