Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals
Patent
1995-05-10
1998-11-10
Green, Lora M.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
435 6, 435 71, 435 79, 435 792, 435962, 435967, 436501, 436524, 436527, 436528, 436531, 436532, 436533, 436534, 436161, 436802, 436808, 436815, G01N 3353
Patent
active
058343180
ABSTRACT:
Ligands that interact with a target can be more easily identified if false positive interactions (either specific or non-specific) from the detecting system are differentiated from the target-specific interaction. An improved method of identifying peptides which bind with a target protein is presented. The steps are: binding a random library of peptides to a support material, allowing detection reagents to contact the peptides and the support material then identifying these interactions, then allowing the target protein to selectively bind to the peptides, allowing detection reagents to contact the bound target protein, and characterizing the peptide bound to the identified support material. Interaction of a ligand or the support material with the detection reagents will cause a distinct color change which distinguishes those ligands which selectively bind to target protein. The characterized peptide can then be used in affinity purification of the target protein. In one embodiment, automation of the assay is demonstrated by flowing all immunoreagents through the beads in a column format ensuring highly efficient washing. In the preferred embodiment, a resin for peptide synthesis which is hydrophilic, contains spacers and may exhibit less nonspecific background than other resins permits synthesis and direct evaluation of combinatorial peptide libraries for binding to target proteins is utilized. Examples for the use of this new resin and methodology for identifying peptide-ligands for purification of proteins are presented.
REFERENCES:
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Vagner, J., et al., "Novel methodology for differentiation of `surface` and `interior` areas of polyoxyethylene-polystyrene (POE-PS) supports: application to library screening procedures," in Innovative Perspectives in Solid Phase Synthesis (R. Epton, ed., 1994), pp. 347-352.
Lam, K.S., et al. "Application of a dual color selection scheme in the screening of a random combinatorial peptide library," J. Immunol. Meth. 180: 219-223 (1995).
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Bayer Corporation
Beck Michael A.
Giblin James A.
Green Lora M.
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