Patent US4629175A successfully refreshed from Google Patents.
US5225533A
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US
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General method for producing and selecting peptides with specific properties
Assignee: University of California, San Diego
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Filing: 1991-03-22
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Granted: 1993-07-06
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Expires: 2011-03-22
Abstract
Abstract A method to obtain selected individual peptides or families thereof which have a target property and optionally to determine the amino acid sequence of a selected peptide or peptides to permit synthesis in practical quantities is disclosed. In general outline, the method of the invention comprises synthesizing a mixture of randomly or deliberately generated peptides using standard synthesis techniques, but adjusting the individual concentrations of the components of a mixture of sequentially added amino acids according to the coupling constants for each amino acid/amino acid coupling. The subgroup of peptides having the target property can then be selected, and either each peptide isolated and sequenced, or analysis performed on the mixture to permit its composition to be reproduced. Also included in the invention is an efficient method to determine the relevant coupling constants.
Inventors
- 👤 William J. Rutter
- 👤 Daniel V. Santi
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 02:45
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; C; C07; C07K; C07K1/00; C07K1/04; C07K1/047; C07K1/14; C07K1/36
Claims Analysis
Total Claims: 41
Claim 1
Independent Claim
92 words
A method of preparing a mixture of distinct, unique and different peptides in the same reaction vessel, which mixture contains each peptide in retrievable and analyzable amounts, comprising: combining and reacting activated amino acids with an acceptor amino acid or peptide wherein said activated amino acids are provided in concentrations relative to each other based on the relative coupling constants so that the mixture of the peptides resulting from the reaction contains each of the peptides in predictable and defined amounts sufficient for each of the peptides to be retrieved and analyzed.
Claim 2
Independent Claim
92 words
A method of preparing a mixture of distinct, unique and different peptides in the same reaction vessel, which mixture contains each peptide in retrievable and analyzable amounts, comprising: combining and reacting activated amino acids with an acceptor amino acid or peptide wherein said activated amino acids are provided in concentrations relative to each other based on the relative coupling constants so that the mixture of the peptides resulting from the reaction contains each of the peptides in predictable and defined amounts sufficient for each of the peptides to be retrieved and analyzed.
Claim 3
Dependent Claim
28 words
The method of preparing a mixture of peptides as claimed in claim 1, further comprising: retrieving at least one peptide from the mixture having a selected target property.
Claim 4
Dependent Claim
31 words
The method of preparing a mixture of peptides as claimed in claim 2, further comprising: analyzing the peptide having the selected target property in order to determine its amino acid sequence.
Claim 5
Dependent Claim
37 words
The method of preparing a mixture of peptides as claimed in claim 1 wherein the combining and reacting is carried out using an acceptor amino acid or peptide derivatized to a solid support through its carboxy terminus.
Claim 6
Dependent Claim
30 words
The method of preparing a mixture of peptides as claimed in claim 4, wherein the method employs a multiplicity of acceptor amino acids or peptides derivatized to the solid support.
Claim 7
Dependent Claim
30 words
The method of preparing a mixture of peptides as claimed in claim 4, wherein a multiplicity of acceptor amino acids are derivatized to a solid support through their carboxy terminus.
Claim 8
Dependent Claim
29 words
The method of preparing a mixture of peptides as claimed in claim 1, wherein the concentration of any given activated amino acid is inversely proportional to its coupling constant.
Claim 9
Independent Claim
120 words
A method of obtaining a peptide or mixture of peptides of a specified target property, which method comprises the steps of: reacting, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein, based on their relative coupling constants to thereby synthesize a mixture of distinct, unique and different candidate peptides in predictable and defined amounts in the same reaction vessel, which mixture contains at least about fifty candidate peptides with each of the candidate peptides being present in retrievable and analyzable amounts; and selecting from among the mixture of candidate peptides one or more peptides having a desired target property and separating such away from those not having the target property.
Claim 10
Independent Claim
120 words
A method of obtaining a peptide or mixture of peptides of a specified target property, which method comprises the steps of: reacting, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein, based on their relative coupling constants to thereby synthesize a mixture of distinct, unique and different candidate peptides in predictable and defined amounts in the same reaction vessel, which mixture contains at least about fifty candidate peptides with each of the candidate peptides being present in retrievable and analyzable amounts; and selecting from among the mixture of candidate peptides one or more peptides having a desired target property and separating such away from those not having the target property.
Claim 11
Dependent Claim
33 words
The method of obtaining a peptide as claimed in claim 8, wherein the selection process utilizes a combination of diode ray detection liquid chromatography, mixture peptide sequencing, mass spectroscopy and amino acid analysis.
Claim 12
Dependent Claim
19 words
The method as claimed in claim 8, wherein the target property is binding specifically to a peptide or protein.
Claim 13
Dependent Claim
25 words
The method as claimed in claim 8, wherein the protein is selected from an immunoglobin, a receptor, an enzyme, an antigen, and a receptor-binding liquid.
Claim 14
Dependent Claim
18 words
The method as claimed in claim 8, wherein the target property is binding specifically to DNA or RNA.
Claim 15
Dependent Claim
18 words
The method as claimed in claim 8, wherein the target property is binding to a carbohydrate or glycoprotein.
Claim 16
Dependent Claim
18 words
The method as claimed in claim 8, wherein the target property is transport or passage through a membrane.
Claim 17
Dependent Claim
16 words
The method as claimed in claim 8, wherein the target property is retention by a membrane.
Claim 18
Dependent Claim
22 words
The method as claimed in claim 8, wherein the target property is utilization as a substrate or inhibitor for a designated enzyme.
Claim 19
Dependent Claim
27 words
The method as claimed in claim 8, wherein the target property is selected from the group consisting of physical characteristics, freezing point, molecular size, and solubility profile.
Claim 20
Dependent Claim
93 words
The method as claimed in claim 8, wherein the step of synthesizing the peptide is carried out by a process comprising the steps of: determining the relative coupling constants of a set of activated amino acids to be reacted; and combining and reacting the activated amino acids with an acceptor amino acid or peptide wherein the activated amino acids are provided in concentrations relative to each other based on the relative coupling constants so that the mixture of peptides resulting from the reaction contains each of the peptides in retrievable and analyzable amounts.
Claim 21
Dependent Claim
35 words
The method as claimed in claim 18, wherein the combining and reacting is carried out in a step wherein the acceptor amino acid or peptide is derivatized to a solid support through its carboxy terminus.
Claim 22
Dependent Claim
32 words
The method as claimed in claim 19, wherein the concentrations of the activated amino acids relative to each other are inversely proportional to the respective coupling constants of the activated amino acids.
Claim 23
Independent Claim
144 words
A method of preparing a mixture of peptides containing a recoverable amount of a peptide having a specified target property, comprising: reacting, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein, based on their relative coupling constants to thereby synthesize a mixture of distinct, unique and different candidate peptides in predictable and defined amounts in the same reaction vessel in such a manner that the mixture will contain at least about fifty candidate peptides with each being present in a retrievable and analyzable amount; treating the mixture of peptides under conditions wherein the peptides of the specified target property is placed in condition to be separated from the remaining peptides; separating the peptides with the specified target property from peptides not possessing the target property; and recovering the peptides with the specified target property.
Claim 24
Independent Claim
144 words
A method of preparing a mixture of peptides containing a recoverable amount of a peptide having a specified target property, comprising: reacting, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein, based on their relative coupling constants to thereby synthesize a mixture of distinct, unique and different candidate peptides in predictable and defined amounts in the same reaction vessel in such a manner that the mixture will contain at least about fifty candidate peptides with each being present in a retrievable and analyzable amount; treating the mixture of peptides under conditions wherein the peptides of the specified target property is placed in condition to be separated from the remaining peptides; separating the peptides with the specified target property from peptides not possessing the target property; and recovering the peptides with the specified target property.
Claim 25
Dependent Claim
35 words
The method as claimed in claim 21, wherein the conditions include contacting the mixture with a reagent which binds specifically to the desired peptide with the target property; and separating the bound from unbound peptides.
Claim 26
Dependent Claim
15 words
The method as claimed in claim 22, wherein the reagent is a protein or peptide.
Claim 27
Dependent Claim
31 words
The method as claimed in claim 23, wherein the reagent protein or peptide is selected from the group consisting of immunoglobin, a receptor, an enzyme, an antigen, and a receptor-binding ligand.
Claim 28
Dependent Claim
20 words
The method as claimed in claim 22, wherein the reagent is selected from the group consisting of DNA and RNA.
Claim 29
Dependent Claim
22 words
The method as claimed in claim 22, wherein the reagent is selected from the group consisting of a carbohydrate and a glycoprotein.
Claim 30
Dependent Claim
29 words
The method as claimed in claim 22, wherein the conditions including contacting the mixture with a membrane through which only the desired peptides having the target property are transported.
Claim 31
Dependent Claim
29 words
The method as claimed in claim 22, wherein the conditions include contacting the mixture with a membrane through which only the desired peptides having the target property are retained.
Claim 32
Dependent Claim
24 words
The method as claimed in claim 22, wherein the conditions include contacting the mixture with a means for separating the proteins according to size.
Claim 33
Dependent Claim
17 words
The method as claimed in claim 22, wherein the conditions include lowering the temperature of the mixture.
Claim 34
Dependent Claim
24 words
The method as claimed in claim 22, wherein the conditions include means for separating according to differences in the physical characteristics of the proteins.
Claim 35
Dependent Claim
18 words
The method as claimed in claim 22, further comprising: determining the amino acid sequence of the desired peptide.
Claim 36
Independent Claim
137 words
The method of preparing a mixture of distinct, unique and different peptides in predictable and defined amounts in the same reaction vessel, which mixture contains a peptide of a desired amino acid sequence and a specified target property, comprising: (1) treating, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein to obtain a set of peptides the acceptor amino acids being added in a concentration based on relative coupling constants so that each such peptide is present in the set in retrievable amounts; (2) deprotecting the peptides; (3) treating the deprotected peptides with an excess of a single N-blocked, carboxy-activated amino acid under conditions with effect conjugation to obtain a set of peptides such that each peptide is present in retrievable amounts; and (4) deprotecting the peptides.
Claim 37
Independent Claim
137 words
The method of preparing a mixture of distinct, unique and different peptides in predictable and defined amounts in the same reaction vessel, which mixture contains a peptide of a desired amino acid sequence and a specified target property, comprising: (1) treating, under conditions which effect conjugation, a mixture of N-blocked, carboxy-activated amino acids with an acceptor amino acid or protein to obtain a set of peptides the acceptor amino acids being added in a concentration based on relative coupling constants so that each such peptide is present in the set in retrievable amounts; (2) deprotecting the peptides; (3) treating the deprotected peptides with an excess of a single N-blocked, carboxy-activated amino acid under conditions with effect conjugation to obtain a set of peptides such that each peptide is present in retrievable amounts; and (4) deprotecting the peptides.
Claim 38
Dependent Claim
27 words
The method as claimed in claim 3, which further comprises repeating steps (1) and (2) after the completion of step (2) but prior to beginning step (3).
Claim 39
Dependent Claim
21 words
The method as claimed in claim 3, which further includes repeating steps (3) and (4) after the completion of step (4).
Claim 40
Dependent Claim
25 words
The method as claimed in claim 33, which further comprises selecting from among the resulting peptides one or more peptides having the desired target property.
Claim 41
Dependent Claim
37 words
The method as claimed in claim 33, wherein the concentration of each N-blocked, carboxy-activated amino acid is adjusted to yield the desired protein according to its coupling constant with respect to the acceptor amino acid or peptide.
References Cited (Backward Citations)
WO1986000991A1
Cited
US5010175A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1996-07-05 | 1997-01-04 | 1997-07-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-07-05 | 2001-01-04 | 2001-07-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-07-05 | 2005-01-04 | 2005-07-06 | $7700.00 | Window Closed / Lapsed |