US5789208A
Expired / Lapsed
US
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Polyclonal antibody libraries
Assignee: Boston University
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Filing: 1997-02-19
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Granted: 1998-08-04
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Expires: 2017-02-19
Abstract
Abstract The invention is directed to methods for the creation and use of libraries of proteins which comprise polyclonal antibodies to a common antigen or group of antigens, receptor proteins with related variable regions, or other immune related proteins with variable regions. These polyclonal antibody libraries can be used to treat or prevent diseases and disorders including neoplasia such as cancer and other malignancies, parasitic infections, bacterial infections, viral infections and disorders such as genetic defects and deficiencies. Protein libraries may be patient-specific, disease-specific or both patient- and disease-specific. Libraries can also be used to detect a disease or disorder in a patient either by direct imaging or through the use of a diagnostic kit. The invention further includes novel cloning methods for the creation and transfer of nucleic acid sequences encoding protein variable regions and novel cloning vectors.
Inventors
- 👤 Jacqueline Sharon
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 16:21
CPC Classifications
C; C40; C40B; C40B40/00; C40B40/02; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/68; A61K47/6835; A61K47/6851; C07; C07K; C07K16/00; C07K16/18; C07K16/28; C07K16/30; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1034; C12N15/1037; A61K39/00; A61K2039/505; A61K38/00; C07K2317/00; C07K2317/20; C07K2317/24; C07K2317/50; C07K2317/55
Claims Analysis
Total Claims: 27
Claim 1
Independent Claim
235 words
A method for preparing a first library of expression vectors comprising paired nucleic acid fragments, said paired nucleic acid fragments being suitable for transfer to prepare a second library of different expression vectors, comprising the steps of: (a) obtaining a plurality of nucleic acid fragments wherein each nucleic acid fragment encodes a variable region or part thereof of a protein which contains said variable region; (b) producing a plurality of nucleic acid segments, wherein each segment comprises a pair of different nucleic acid fragments selected from said plurality of nucleic acid fragments and linked in opposite and divergent transcriptional orientations; (c) cloning said plurality of nucleic acid segments into first circular vectors to produce a library of vector molecules; (d) inserting a first cassette between the two nucleic acid fragments of each nucleic acid segment to produce a nucleic acid segment comprising said first cassette in a first library of expression vector molecules, wherein said first cassette contains at least one promoter element permitting expression of said nucleic acid fragments in a first host cell; (e) introducing said first library of expression vector molecules into the first host cells under conditions and time sufficient to express a first library of polypeptides, wherein each of the polypeptides contains a sequence encoded by one of said pairs of linked nucleic acid fragments; and (f) isolating said first library of expression vector molecules from said first host cells.
Claim 2
Independent Claim
235 words
A method for preparing a first library of expression vectors comprising paired nucleic acid fragments, said paired nucleic acid fragments being suitable for transfer to prepare a second library of different expression vectors, comprising the steps of: (a) obtaining a plurality of nucleic acid fragments wherein each nucleic acid fragment encodes a variable region or part thereof of a protein which contains said variable region; (b) producing a plurality of nucleic acid segments, wherein each segment comprises a pair of different nucleic acid fragments selected from said plurality of nucleic acid fragments and linked in opposite and divergent transcriptional orientations; (c) cloning said plurality of nucleic acid segments into first circular vectors to produce a library of vector molecules; (d) inserting a first cassette between the two nucleic acid fragments of each nucleic acid segment to produce a nucleic acid segment comprising said first cassette in a first library of expression vector molecules, wherein said first cassette contains at least one promoter element permitting expression of said nucleic acid fragments in a first host cell; (e) introducing said first library of expression vector molecules into the first host cells under conditions and time sufficient to express a first library of polypeptides, wherein each of the polypeptides contains a sequence encoded by one of said pairs of linked nucleic acid fragments; and (f) isolating said first library of expression vector molecules from said first host cells.
Claim 3
Dependent Claim
50 words
The method of claim 1, which further comprises isolating the plurality of segments comprising said first cassette from said first library of expression vectors molecules, and inserting said plurality of segments comprising said first cassette into a plurality of second vectors to produce a second library of expression vector molecules.
Claim 4
Dependent Claim
51 words
The method of claim 1, which further comprises exchanging said first cassette from an expression vector of said isolated first library of expression vector molecules with a second cassette, wherein said second cassette contains at least one promoter element permitting expression of said nucleic acid fragments in a second host cell.
Claim 5
Dependent Claim
50 words
The method of claim 2, which further comprises exchanging said first cassette from an expression vector of said second library of expression vector molecules with a second cassette, wherein said second cassette contains at least one promoter element permitting expression of said nucleic acid fragments in a second host cell.
Claim 6
Dependent Claim
22 words
The method of claim 2, wherein said second library of expression vector molecules is introduced into a plurality of second host cells.
Claim 7
Dependent Claim
13 words
The method of claim 4, wherein the second vector is a circular vector.
Claim 8
Dependent Claim
23 words
The method of claim 5, wherein said first host cell is a prokaryotic cell and said second host cell is a eukaryotic cell.
Claim 9
Dependent Claim
24 words
The method of claim 5, wherein said first host cell is a eukaryotic cell and said second host cell is a second eukaryotic cell.
Claim 10
Dependent Claim
24 words
The method of claim 5, wherein said first host cell is a prokaryotic cell and said second host cell is a second prokaryotic cell.
Claim 11
Dependent Claim
23 words
The method of claim 5, wherein said first host cell is a eukaryotic cell and said second host cell is a prokaryotic cell.
Claim 12
Dependent Claim
15 words
The method of claim 1, wherein said protein is a secreted protein or receptor protein.
Claim 13
Dependent Claim
24 words
The method of claim 11, wherein the secreted or receptor protein is selected from the group consisting of an antibody and a T-cell receptor.
Claim 14
Dependent Claim
48 words
The method of claim 1, wherein one nucleic acid fragment of each segment of said plurality of nucleic acid segments encodes a heavy chain variable region of an antibody and the other nucleic acid fragment of the segment encodes a light chain variable region of the same antibody.
Claim 15
Dependent Claim
48 words
The method of claim 1 wherein said nucleic acid fragments comprise a polyclonal population encoding variable regions or portions thereof of proteins selected from the group consisting of antibodies, T-cell receptors, B-cell receptors, natural killer cell receptors, macrophage receptors and combinations thereof derived from a polyclonal cell population.
Claim 16
Dependent Claim
15 words
The method of claim 14, wherein the proteins are selected from antibodies or T-cell receptors.
Claim 17
Dependent Claim
16 words
The method of claim 1 wherein the first cassette contains two promoters in opposite transcriptional orientations.
Claim 18
Dependent Claim
16 words
The method of claim 3 wherein the second cassette contains two promoters in opposite transcriptional orientations.
Claim 19
Dependent Claim
50 words
The method of claim 1, wherein said first cassette further comprises a nucleic acid sequence encoding a leader sequence of a secreted protein or membrane protein, and said nucleic acid sequence encoding said leader sequence is operably linked to a nucleic acid fragment of said pair of nucleic acid fragments.
Claim 20
Dependent Claim
50 words
The method of claim 3, wherein said first cassette further comprises a nucleic acid sequence encoding a leader sequence of a secreted protein or membrane protein, and said nucleic acid sequence encoding said leader sequence is operably linked to a nucleic acid fragment of said pair of nucleic acid fragments.
Claim 21
Dependent Claim
21 words
The method of claim 1, wherein said first circular vector is a prokaryotic or eukaryotic surface display vector or expression vector.
Claim 22
Dependent Claim
13 words
The method of claim 2 wherein said second vector is a circular vector.
Claim 23
Dependent Claim
21 words
The method of claim 21, wherein said second circular vector is a prokaryotic or eukaryotic surface display vector or expression vector.
Claim 24
Dependent Claim
13 words
The method of claim 1 wherein said polypeptides of step (e) are antibodies.
Claim 25
Dependent Claim
11 words
The method of claim 23, wherein said antibodies are chimeric antibodies.
Claim 26
Dependent Claim
17 words
The method of claim 24, wherein said chimeric antibodies have mouse variable regions and human constant regions.
Claim 27
Dependent Claim
42 words
The method of claim 1, wherein step (f) includes a preliminary screening step to select desired expression vector molecules and thereby reduce the number of expression vector molecules used in preparation of said second library from those used in said first library.
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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) | 2001-08-03 | 2002-02-02 | 2002-08-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2005-08-03 | 2006-02-02 | 2006-08-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2009-08-03 | 2010-02-02 | 2010-08-04 | $7700.00 | Window Closed / Lapsed |