US5010175A
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US
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General method for producing and selecting peptides with specific properties
Assignee: CALIFORNIA 2199 ADDISON STREET BERKELEY CA 94720, University of, Regents of
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Filing: 1988-05-02
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Granted: 1991-04-23
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Expires: 2008-05-02
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
2026-10-23 03:44
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/04; C07K1/047; C07K1/14; C07K1/36; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68
Claims Analysis
Total Claims: 47
Claim 1
Independent Claim
100 words
A method of preparing in the same reaction vessel a mixture of peptides of distinct, unique and different sequences which mixture contains each peptide in retrievable and analyzable amounts and in substantially equal molar 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
100 words
A method of preparing in the same reaction vessel a mixture of peptides of distinct, unique and different sequences which mixture contains each peptide in retrievable and analyzable amounts and in substantially equal molar 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
26 words
The method of preparing a mixture of peptides as claimed in claim 2, wherein each peptide produced in the mixture is produced in equal molar amounts.
Claim 7
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 8
Dependent Claim
45 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 and further wherein each peptide in the mixture is produced in substantially equal molar amounts.
Claim 9
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 10
Independent Claim
86 words
A method of obtaining a peptide or mixture of peptides of a specified target property, which method comprises the steps of: providing in the same vessel a mixture of candidate peptides which mixture contains at least about fifty candidate peptides with each of the candidate peptides being present in retrievable and analyzable amounts and in substantially equal molar amounts; and selecting from among the candidate peptides one or more peptides having a desired target property and separating such away from those not having the target property.
Claim 11
Independent Claim
86 words
A method of obtaining a peptide or mixture of peptides of a specified target property, which method comprises the steps of: providing in the same vessel a mixture of candidate peptides which mixture contains at least about fifty candidate peptides with each of the candidate peptides being present in retrievable and analyzable amounts and in substantially equal molar amounts; and selecting from among the candidate peptides one or more peptides having a desired target property and separating such away from those not having the target property.
Claim 12
Dependent Claim
33 words
The method of obtaining a peptide as claimed in claim 9, wherein the selection process utilizes a combination of diode ray detection liquid chromatography, mixture peptide sequencing, mass spectroscopy and amino acid analysis.
Claim 13
Dependent Claim
19 words
The method as claimed in claim 9, wherein the target property is binding specifically to a peptide or protein.
Claim 14
Dependent Claim
25 words
The method as claimed in claim 9, wherein the protein is selected from an immunoglobin, a receptor, an enzyme, an antigen, and a receptor-binding ligand.
Claim 15
Dependent Claim
18 words
The method as claimed in claim 9, wherein the target property is binding specifically to DNA or RNA.
Claim 16
Dependent Claim
18 words
The method as claimed in claim 9, wherein the target property is binding to a carbohydrate or glycoprotein.
Claim 17
Dependent Claim
18 words
The method as claimed in claim 9, wherein the target property is transport or passage through a membrane.
Claim 18
Dependent Claim
16 words
The method as claimed in claim 9, wherein the target property si retention by a membrane.
Claim 19
Dependent Claim
22 words
The method as claimed in claim 9, wherein the target property is utilization as a substrate or inhibitor for a designated enzyme.
Claim 20
Dependent Claim
28 words
The method as claimed in claim 9, wherein the target property is a physical characteristics selected from the group consisting of freezing point, molecular size, and solubility profile.
Claim 21
Dependent Claim
93 words
The method as claimed in claim 9, wherein the strep 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 substantially equal molar amounts.
Claim 22
Dependent Claim
35 words
The method as claimed in claim 19, wherein the combining and reacting is carried out in a step wherein the accepto amino acid or peptide is derivatized to a solid support through its carboxy terminus.
Claim 23
Dependent Claim
32 words
The method as claimed in claim 20, 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 24
Independent Claim
105 words
A method of preparing a mixture of peptides containing a recoverable amount of a peptide having a specified target property, comprising: synthesizing a mixture of at least about 50 candidate peptides in such a manner that the mixture will contain each peptide in a retrievable and analyzable amount and in substantially equal molar amounts; treating the mixture containing the 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
Independent Claim
105 words
A method of preparing a mixture of peptides containing a recoverable amount of a peptide having a specified target property, comprising: synthesizing a mixture of at least about 50 candidate peptides in such a manner that the mixture will contain each peptide in a retrievable and analyzable amount and in substantially equal molar amounts; treating the mixture containing the 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 26
Dependent Claim
35 words
The method as claimed in claim 22, 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 27
Dependent Claim
15 words
The method as claimed in claim 23, wherein the reagent is a protein or peptide.
Claim 28
Dependent Claim
30 words
The method as claimed in claim 24, wherein the protein or peptide is selected from the group consisting of immunoglobin, a receptor, an enzyme, an antigen, and a receptor-binding liqand.
Claim 29
Dependent Claim
20 words
The method as claimed in claim 23, wherein the reagent is selected from the group consisting of DNA and RNA.
Claim 30
Dependent Claim
22 words
The method as claimed in claim 23, wherein the reagent is selected from the group consisting of a carbohydrate and a glycoprotein.
Claim 31
Dependent Claim
29 words
The method as claimed in claim 23, wherein the conditions include contacting the mixture with a membrane through which only the desired peptides having the target property are transported.
Claim 32
Dependent Claim
29 words
The method as claimed in claim 23, wherein the conditions include contacting the mixture with a membrane through which only the desired peptides having the target property are retained.
Claim 33
Dependent Claim
24 words
The method as claimed in claim 23, wherein the conditions include contacting the mixture with a means for separating the proteins according to size.
Claim 34
Dependent Claim
17 words
The method as claimed in claim 23, wherein the conditions include lowering the temperature of the mixture.
Claim 35
Dependent Claim
24 words
The method as claimed in claim 23, wherein the conditions include means for separating according to differences in the physical characteristics of the proteins.
Claim 36
Dependent Claim
18 words
The method as claimed in claim 23, further comprising: determining the amino acid sequence of the desired peptide.
Claim 37
Dependent Claim
15 words
The method as claimed in claim 23, further comprising: synthesizing a mixture of candidate peptides.
Claim 38
Independent Claim
144 words
The method of preparing a mixture of peptides of distinct, unique and different sequences containing 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 first-step peptides the acceptor amino acids being added in a concentration based on the relative coupling constants so that each first step peptide is present in the set in retrievable amounts and in substantially equal molar amounts; (2) deprotecting the first-step peptides; (3) treating the deprotected peptides with an excess of a single N-blocked, carboxy-activated amino acid under conditions which effect conjugation to obtain a set of second-step peptides such that each peptide is present in retrievable amounts and in substantially equal molar amounts; (4) deprotecting the second-step peptides.
Claim 39
Independent Claim
144 words
The method of preparing a mixture of peptides of distinct, unique and different sequences containing 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 first-step peptides the acceptor amino acids being added in a concentration based on the relative coupling constants so that each first step peptide is present in the set in retrievable amounts and in substantially equal molar amounts; (2) deprotecting the first-step peptides; (3) treating the deprotected peptides with an excess of a single N-blocked, carboxy-activated amino acid under conditions which effect conjugation to obtain a set of second-step peptides such that each peptide is present in retrievable amounts and in substantially equal molar amounts; (4) deprotecting the second-step peptides.
Claim 40
Dependent Claim
21 words
The method as claimed in claim 35, which further comprises repeating steps (1) and (2) with respect to said first-step peptides.
Claim 41
Dependent Claim
21 words
The method as claimed in claim 35, which further comprises repeating steps (3) and (4) with respect to said second-step peptides.
Claim 42
Dependent Claim
26 words
The method as claimed in claim 35, which further comprises selecting from among the resulting candidate peptides one or more peptides having the desired target property.
Claim 43
Dependent Claim
30 words
The method as claimed in claim 35, wherein the concentration of each N-blocked, carboxy-activated amino acid is adjusted to yield substantially equal molar amounts of each of the first-step peptides.
Claim 44
Independent Claim
74 words
A method of determining the sequence of a peptide containing at least fifty peptide in a mixture of peptides wherein the mixture of peptides resulted from a reaction of distinct, unique and different sequences or set of reactions which produced each peptide in the mixture in substantially equal molar amounts and subjecting the mixture to a battery of analytical techniques to obtain analytical data which is manipulated to determine the sequence of individual peptides.
Claim 45
Independent Claim
74 words
A method of determining the sequence of a peptide containing at least fifty peptide in a mixture of peptides wherein the mixture of peptides resulted from a reaction of distinct, unique and different sequences or set of reactions which produced each peptide in the mixture in substantially equal molar amounts and subjecting the mixture to a battery of analytical techniques to obtain analytical data which is manipulated to determine the sequence of individual peptides.
Claim 46
Dependent Claim
17 words
The method as claimed in claim 40, wherein the analytical techniques include HPLC with diode ray detection.
Claim 47
Dependent Claim
14 words
The method as claimed in claim 40, wherein the analytical techniques include mass spectroscopy.
References Cited (Backward Citations)
No citation records indexed.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1994-04-22 | 1994-10-22 | 1995-04-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-04-22 | 1998-10-22 | 1999-04-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-04-22 | 2002-10-22 | 2003-04-23 | $7700.00 | Window Closed / Lapsed |