US5795747A
Expired / Lapsed
US
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Process for manufacturing new biopolymers
Assignee: Evotec OAI AG
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Filing: 1995-06-26
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Granted: 1998-08-18
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Expires: 2015-06-26
Abstract
Abstract The process of generating new biopolymers with improved characteristics, assisted by polymerases, is characterized in that at least one cycle of the steps described below, will be performed in a series of parallel preparations which are to be compared. Sequences of nucleic acids or, a mixture of similar sequences of nucleic acids in the distribution of mutants of a quasi-species, are subjected, in the region of the error threshold, to limited mutagenesis. These mixtures are replicated under simultaneous conditions and/or in succession. The mixtures of the so product nucleic acids are compartmentalized by segregation, and thereafter selected by a selection system which reflects the characteristics of interest of the nucleic acid sequence itself or, indirectly, via its translation product.
Inventors
- 👤 Karsten Henco
- 👤 Manfred Eigen
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 05:26
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1079; C12N15/1058; C12Q; C12Q1/00; C12Q1/68; C12Q1/6811
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
153 words
A process of preparing new biopolymers with improved characteristics starting with nucleic acids comprising the steps of: a) subjecting to limited mutagenesis in the range of the error threshold at least one sample of identical nucleic acid sequences or similar sequences of nucleic acids in the mutant distribution of a quasi-species to effect a mixture of nucleic acid sequences; b) replicating, under less stringent mutagenic pressure than step (a), said mixture of nucleic acid sequences in said at least one sample to effect mutant distributions; c) separating said mutant distributions into secondary samples; d) repeating steps a)-c) in at least one cycle; e) performing steps a) and b); f) selecting at least one of said secondary samples based on phenotypic characteristics of the mutuant distributions in the secondary sample; and g) isolating from said selected secondary sample a mutagenized nucleic acid sequence based on its characteristics or the characteristics of its translation product.
Claim 2
Dependent Claim
153 words
A process of preparing new biopolymers with improved characteristics starting with nucleic acids comprising the steps of: a) subjecting to limited mutagenesis in the range of the error threshold at least one sample of identical nucleic acid sequences or similar sequences of nucleic acids in the mutant distribution of a quasi-species to effect a mixture of nucleic acid sequences; b) replicating, under less stringent mutagenic pressure than step (a), said mixture of nucleic acid sequences in said at least one sample to effect mutant distributions; c) separating said mutant distributions into secondary samples; d) repeating steps a)-c) in at least one cycle; e) performing steps a) and b); f) selecting at least one of said secondary samples based on phenotypic characteristics of the mutuant distributions in the secondary sample; and g) isolating from said selected secondary sample a mutagenized nucleic acid sequence based on its characteristics or the characteristics of its translation product.
Claim 3
Dependent Claim
20 words
The process according to claim 1, wherein steps a)-f) are repeated in at least one cycle before performing step g).
Claim 4
Dependent Claim
26 words
The process according to claim 1, wherein said limited mutagenesis is performed by (i) contacting the sample with a mutagenic agent or (ii) irradiating the sample.
Claim 5
Dependent Claim
53 words
The process according to claim 3, wherein the sample is contacted with an unbalanced dose of a mutagenic agent and wherein said mutagenic agent is selected from the group consisting of a purine and a pyrimidine nucleotide base, a base analog thereof, and a combination of the nucleotide base and nucleotide base analog.
Claim 6
Dependent Claim
69 words
The process according to claim 1, wherein the distribution of the nucleic acid sequences is guided in the range of the error threshold through the value landscape in the direction of a desired optimum (i) by adjusting the accuracy of replication used as a variable or by gradating the accuracy in consecutive steps in the range of the error threshold followed by (ii) >>10 times more exact replicative amplification.
Claim 7
Dependent Claim
22 words
The process according to claim 1, wherein the limited mutagenesis involves exposing different sections of the nucleic acids to different mutagenic pressures.
Claim 8
Dependent Claim
30 words
The process according to claim 1, wherein nucleic acid sequences in a secondary sample selected by phenotype are classified simultaneously or later on and further alone or in a mixture.
Claim 9
Dependent Claim
31 words
The process according to claim 1, wherein the mutagenized nucleic acid is isolated based on its translation product in which said mutagenized nucleic acid and said translation product are directly coupled.
Claim 10
Dependent Claim
29 words
The process according to claim 1, wherein said isolation is carried out using polysomes that bind, in a coupled form, said translation product and its coding mutagenized nucleic acid.
Claim 11
Dependent Claim
26 words
The process according to claim 1, wherein the selecting step is controlled, cycle by cycle, by varying the stringency of limited mutagenesis conditions and selecting conditions.
Claim 12
Dependent Claim
13 words
The process according to claim 1, conducted simultaneously on two or more samples.
Claim 13
Dependent Claim
12 words
The process according to claim 11, involving at least 10 3 samples.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2001-08-17 | 2002-02-16 | 2002-08-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-08-17 | 2006-02-16 | 2006-08-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-08-17 | 2010-02-16 | 2010-08-18 | $7700.00 | Window Closed / Lapsed |