Method of screening a peptide library

Claim 1

A method of screening a library of peptides and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library comprises a multiplicity of species of solid phase supports having a linker, by which analyzable quantities of deprotected peptides are covalently attached to the support, wherein the sequences of said peptides are not pre-determined and a single species of peptide is attached to each support; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule, wherein said identified supports represent at most about 1 part per 1,000 parts of the supports of the library; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 2

A method of screening a library of peptides and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library comprises a multiplicity of species of solid phase supports having a linker, by which analyzable quantities of deprotected peptides are covalently attached to the support, wherein the sequences of said peptides are not pre-determined and a single species of peptide is attached to each support; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule, wherein said identified supports represent at most about 1 part per 1,000 parts of the supports of the library; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 6

A method of screening a peptide library and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library is comprised of a multiplicity of solid phase supports having a linker, wherein a single peptide species is covalently attached by the linker to each solid phase support, and in which the library is prepared by a method comprising the steps of: (i) providing at least two aliquots of solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunit to substantially all the reactive sites of the solid phase supports; (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports whereby an N.sup.α reactive site is provided; and after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that the peptides remain covalently attached to the solid phase supports; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule, wherein said identified supports represent at most about 1 part per 1,000 parts of the supports of the library; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 7

A method of screening a peptide library and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library is comprised of a multiplicity of solid phase supports having a linker, wherein a single peptide species is covalently attached by the linker to each solid phase support, and in which the library is prepared by a method comprising the steps of: (i) providing at least two aliquots of solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunit to substantially all the reactive sites of the solid phase supports; (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports whereby an N.sup.α reactive site is provided; and after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that the peptides remain covalently attached to the solid phase supports; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule, wherein said identified supports represent at most about 1 part per 1,000 parts of the supports of the library; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 13

A method of screening a library of peptides and determining a sequence of a peptide ligand having a biological activity of interest comprising the steps of: a) subdividing a peptide library into a multiplicity of samples by placing each sample into a corresponding sample of solution, wherein the library comprises a multiplicity of species of solid phase supports having analyzable quantifies of deprotected peptides covalently attached thereto by means of a selectively cleavable linker, a plurality of selectively cleavable linkers or a combination of a non-cleavable linker and a selectively cleavable linker, wherein the sequences of said peptides are not pre-determined and a single species of peptide is covalently attached to each support; b) releasing a portion of the peptides of each sample of supports into the corresponding sample of solution by cleavage of the cleavable linker; c) detecting the biological activity of interest in the samples of solution; d) identifying the solid-phase supports having peptides thereon that are biologically active, wherein said identified supports represent at most about 1 part per 2,000 part of the supports of the library; e) isolating at least one identified support from all other species of the library; and thereafter f) determining the sequence of the peptide species attached to the isolated support.

Claim 14

A method of screening a library of peptides and determining a sequence of a peptide ligand having a biological activity of interest comprising the steps of: a) subdividing a peptide library into a multiplicity of samples by placing each sample into a corresponding sample of solution, wherein the library comprises a multiplicity of species of solid phase supports having analyzable quantifies of deprotected peptides covalently attached thereto by means of a selectively cleavable linker, a plurality of selectively cleavable linkers or a combination of a non-cleavable linker and a selectively cleavable linker, wherein the sequences of said peptides are not pre-determined and a single species of peptide is covalently attached to each support; b) releasing a portion of the peptides of each sample of supports into the corresponding sample of solution by cleavage of the cleavable linker; c) detecting the biological activity of interest in the samples of solution; d) identifying the solid-phase supports having peptides thereon that are biologically active, wherein said identified supports represent at most about 1 part per 2,000 part of the supports of the library; e) isolating at least one identified support from all other species of the library; and thereafter f) determining the sequence of the peptide species attached to the isolated support.

Claim 25

A method of screening a library of peptides and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library is comprised of a multiplicity of solid phase supports having a linker, wherein a single peptide species is attached covalently by the linker to each solid phase support, and in which the library is prepared by a method comprising at least three iterations of the steps of: (i) providing at least two aliquots of solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunit to substantially all the reactive sites of the solid phase supports; (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports whereby an N.sup.α reactive site is provided; (v) repeating steps (i)-(iv) at least two additional times; and (vi) after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that the peptides remain attached to the solid phase supports; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 26

A method of screening a library of peptides and determining a sequence of a peptide ligand for an acceptor molecule of interest comprising the steps of: a) introducing an acceptor molecule to a peptide library, wherein the library is comprised of a multiplicity of solid phase supports having a linker, wherein a single peptide species is attached covalently by the linker to each solid phase support, and in which the library is prepared by a method comprising at least three iterations of the steps of: (i) providing at least two aliquots of solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunit to substantially all the reactive sites of the solid phase supports; (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports whereby an N.sup.α reactive site is provided; (v) repeating steps (i)-(iv) at least two additional times; and (vi) after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that the peptides remain attached to the solid phase supports; b) identifying the solid-phase supports having peptides thereon that bind the acceptor molecule; c) isolating at least one identified support from all other species of the library; and thereafter d) determining the sequence of the peptide species attached to the isolated support.

Claim 30

A method of screening a library of peptides and determining a sequence of a peptide ligand having a biological activity of interest comprising the steps of: a) subdividing a peptide library into a multiplicity of samples by placing each sample into a corresponding sample of solution, wherein a single peptide species is attached covalently to each solid phase support by means of a selectively cleavable linker, a plurality of selectively cleavable linkers or a combination of a non-cleavable linker and a selectively cleavable linker, and in which the library is prepared by a method comprising the steps of: (i) providing at least two aliquots of a solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunits to substantially all the reactive sites of the solid phase supports; and (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports, whereby an N.sup.α reactive site is provided; and, after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that peptides remain linked to the solid phase supports; b) releasing a portion of the peptides of each sample of supports into the corresponding sample of solution by cleavage of the cleavable linker; c) detecting the biological activity of interest in the samples of solution; d) identifying the solid phase supports having peptides thereon that are biologically active, wherein said identified supports represent at most about 1 part per 2,000 part of the supports of the library; e) isolating at least one active support from all other species of the library; and thereafter f) determining the sequence of the peptide species attached to the isolated support.

Claim 31

A method of screening a library of peptides and determining a sequence of a peptide ligand having a biological activity of interest comprising the steps of: a) subdividing a peptide library into a multiplicity of samples by placing each sample into a corresponding sample of solution, wherein a single peptide species is attached covalently to each solid phase support by means of a selectively cleavable linker, a plurality of selectively cleavable linkers or a combination of a non-cleavable linker and a selectively cleavable linker, and in which the library is prepared by a method comprising the steps of: (i) providing at least two aliquots of a solid phase supports, said solid phase supports having an N.sup.α reactive site; (ii) separately introducing a species of subunit of the peptide to each of the aliquots of solid phase supports such that a different species of subunit is introduced into each aliquot, said subunits having a protecting group or a plurality of protecting groups; (iii) completely coupling the subunits to substantially all the reactive sites of the solid phase supports; and (iv) thoroughly mixing and deprotecting the aliquots of solid phase supports, whereby an N.sup.α reactive site is provided; and, after completing all desired executions of steps (i) through (iv), a final step of removing any remaining protecting groups such that peptides remain linked to the solid phase supports; b) releasing a portion of the peptides of each sample of supports into the corresponding sample of solution by cleavage of the cleavable linker; c) detecting the biological activity of interest in the samples of solution; d) identifying the solid phase supports having peptides thereon that are biologically active, wherein said identified supports represent at most about 1 part per 2,000 part of the supports of the library; e) isolating at least one active support from all other species of the library; and thereafter f) determining the sequence of the peptide species attached to the isolated support.