US4879225A
Expired / Lapsed
US
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Enhanced production of antibodies utilizing insolubilized immune complexes
Assignee: Poniard Pharmaceuticals Inc
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Filing: 1987-03-11
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Granted: 1989-11-07
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Expires: 2007-03-11
Abstract
Abstract A method for enhancing production of antibodies through immunization with insolubilized immune complexes is disclosed. Purified antigen or heterogeneous antigen mixtures may be combined with polyclonal or monoclonal antibody and the resultant complex bound to insolubilized protein A to form insolubilized immune complexes. Methods for improving the immunogenicity of a soluble antigen and for producing monoclonal anti-idiotypic antibodies are also disclosed. Monoclonal antibodies that are specific for a distinct, as yet unrecognized epitope may be produced by another disclosed method. Insolubilized immune complexes, comprising antigen and antibody that is either directly linked to Sepharose TM or absorbed onto insolubolized protein A, and immunosorbents, comprising antibody absorbed onto insolubilized protein A, are also disclosed.
Inventors
- 👤 Alton C. Morgan, Jr.
- 👤 Clive S. Woodhouse
- 👤 Robert F. McIntyre
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-25 21:06
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/42; C07K16/4208; C07K16/4241; C07K16/4258; C07K16/4266; A; A61; A61K; A61K39/00; A61K39/385; A61K39/395; A61K39/44; A61K2039/60; A61K2039/6031; A61K2039/6056; A61K2039/62; A61K2039/622; C07K2317/00; C07K2317/30; C07K2317/32
Claims Analysis
Total Claims: 62
Claim 1
Independent Claim
40 words
A method for enhancing production of antibodies, comprising the steps of: absorbing antibody onto insolubilized protein A, thereby forming an immunosorbent; combining an antigen with said immunosorbent, forming an insolubilized immune complex; immunizing an animal with said insolubilized immune complex.
Claim 2
Independent Claim
40 words
A method for enhancing production of antibodies, comprising the steps of: absorbing antibody onto insolubilized protein A, thereby forming an immunosorbent; combining an antigen with said immunosorbent, forming an insolubilized immune complex; immunizing an animal with said insolubilized immune complex.
Claim 3
Dependent Claim
20 words
The method of claim 1 wherein the insolubilized protein A is comprised of protein A bound to an inert matrix.
Claim 4
Dependent Claim
13 words
The method of claim 2 wherein the inert matrix is a polysaccharide matrix.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein the inert polysaccharide matrix is Sepharose®.
Claim 6
Dependent Claim
15 words
The method of claim 2 wherein the inert matrix is in the form of particles.
Claim 7
Dependent Claim
15 words
The method of claim 5 wherein the inert matrix is in the form of particles.
Claim 8
Dependent Claim
23 words
The method of claim 1, comprising, after the step of absorbing the antibody, the additional step of cross-linking the antibody and protein A.
Claim 9
Dependent Claim
22 words
The method of claim 1, comprising, after the step of combining the antigen, the additional step of cross-linking the antibody and antigen.
Claim 10
Dependent Claim
21 words
The method of claim 1 further comprising, after the immunizing step, the step of collecting polyclonal antiserum from said immunized animal.
Claim 11
Dependent Claim
25 words
The method of claim 9 further comprising, after the collecting step, the step of removing non-specific antibodies from the polyclonal antiserum, thereby generating monospecific antiserum.
Claim 12
Dependent Claim
53 words
The method of claim 1 further comprising, after the immunizing step, the steps of: fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing monoclonal antibodies to said antigen; culturing said hybridomas to produce said monoclonal antibodies; and collecting monoclonal antibodies as a product of said hybridomas.
Claim 13
Dependent Claim
12 words
The method of claim 1 wherein the antigen is purified homogeneous antigen.
Claim 14
Dependent Claim
15 words
The method of claim 1 wherein the antigen is a component of a heterogeneous mixture.
Claim 15
Dependent Claim
16 words
The method of claim 1 wherein the antigen is a component of an enriched antigen mixture.
Claim 16
Dependent Claim
18 words
The method of claim 14 wherein the antigen represents at least 1% (w/w) of the enriched antigen mixture.
Claim 17
Dependent Claim
12 words
The method of claim 1 wherein the antigen is a soluble antigen.
Claim 18
Dependent Claim
12 words
The method of claim 1 wherein the antigen is a tumor-associated antigen.
Claim 19
Independent Claim
83 words
A method for enhancing production of specific monoclonal antibodies, comprising the steps of: absorbing antibody onto insolubilized protein A, thereby forming an immunosorbent; combining an antigen with said immunosorbent, forming an insolubilized immune complex; immunizing an animal with said insolubilized immune complex; and fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing monoclonal antibodies to said antigen; culturing said hybridomas to produce said monoclonal antibodies; and collecting said monoclonal antibodies as a product of said hybridomas.
Claim 20
Independent Claim
83 words
A method for enhancing production of specific monoclonal antibodies, comprising the steps of: absorbing antibody onto insolubilized protein A, thereby forming an immunosorbent; combining an antigen with said immunosorbent, forming an insolubilized immune complex; immunizing an animal with said insolubilized immune complex; and fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing monoclonal antibodies to said antigen; culturing said hybridomas to produce said monoclonal antibodies; and collecting said monoclonal antibodies as a product of said hybridomas.
Claim 21
Dependent Claim
20 words
The method of claim 18 wherein the insolubilized protein A is comprised of protein A bound to an inert matrix.
Claim 22
Dependent Claim
13 words
The method of claim 19 wherein the inert matrix is a polysaccharide matrix.
Claim 23
Dependent Claim
12 words
The method of claim 20 wherein the inert polysaccharide matrix is Sepharose®.
Claim 24
Dependent Claim
15 words
The method of claim 19 wherein the inert matrix is in the form of particles.
Claim 25
Dependent Claim
14 words
The method of claim 22 wherein the particles are in the form of beads.
Claim 26
Dependent Claim
23 words
The method of claim 18, comprising, after the step of absorbing the antibody, the additional step of crosslinking the antibody and protein A.
Claim 27
Dependent Claim
22 words
The method of claim 18, comprising, after the step of combining the antigen, the additional step of crosslinking the antibody and antigen.
Claim 28
Dependent Claim
12 words
The method of claim 18 wherein the antigen is purified homogeneous antigen.
Claim 29
Dependent Claim
15 words
The method of claim 18 wherein the antigen is a component of a heterogeneous mixture.
Claim 30
Dependent Claim
16 words
The method of claim 18 wherein the antigen is a component of an enriched antigen mixture.
Claim 31
Dependent Claim
18 words
The method of claim 28 wherein the antigen represents at least 1% (w/w) of the enriched antigen mixture.
Claim 32
Dependent Claim
12 words
The method of claim 18 wherein the antigen is a soluble antigen.
Claim 33
Dependent Claim
12 words
The method of claim 18 wherein the antigen is a tumor-associated antigen.
Claim 34
Independent Claim
123 words
A method for producing monoclonal antibodies specific for an epitope distinct from that recognized by an existing monoclonal antibody, comprising the steps of: absorbing a first monoclonal antibody directed against a first epitope of a polyvalent antigen onto insolubilized protein A, thereby forming an immunosorbent; combining said immunosorbent with said antigen, forming an insolubilized immune complex wherein said first epitope is masked by said first monoclonal antibody; immunizing an animal with said insolubilized immune complex; fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing second monoclonal antibodies to a second epitope of said polyvalent antigen; culturing said hybridomas to produce said second monoclonal antibodies; and collecting said second monoclonal antibodies as a product of said hybridomas.
Claim 35
Independent Claim
123 words
A method for producing monoclonal antibodies specific for an epitope distinct from that recognized by an existing monoclonal antibody, comprising the steps of: absorbing a first monoclonal antibody directed against a first epitope of a polyvalent antigen onto insolubilized protein A, thereby forming an immunosorbent; combining said immunosorbent with said antigen, forming an insolubilized immune complex wherein said first epitope is masked by said first monoclonal antibody; immunizing an animal with said insolubilized immune complex; fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing second monoclonal antibodies to a second epitope of said polyvalent antigen; culturing said hybridomas to produce said second monoclonal antibodies; and collecting said second monoclonal antibodies as a product of said hybridomas.
Claim 36
Dependent Claim
20 words
The method of claim 32 wherein the insolubilized protein A is comprised of protein A bound to an inert matrix.
Claim 37
Dependent Claim
13 words
The method of claim 33 wherein the inert matrix is a polysaccharide matrix.
Claim 38
Dependent Claim
12 words
The method of claim 34 wherein the inert polysaccharide matrix is Sepharose®.
Claim 39
Dependent Claim
15 words
The method of claim 33 wherein the inert matrix is in the form of particles.
Claim 40
Dependent Claim
14 words
The method of claim 36 wherein the particles are in the form of beads.
Claim 41
Dependent Claim
23 words
The method of claim 32, comprising, after the step of absorbing the antibody, the additional step of crosslinking the antibody and protein A.
Claim 42
Dependent Claim
22 words
The method of claim 32, comprising, after the step of combining the antigen, the additional step of crosslinking the antibody and antigen.
Claim 43
Dependent Claim
12 words
The method of claim 32 wherein the antigen is purified homogeneous antigen.
Claim 44
Dependent Claim
15 words
The method of claim 32 wherein the antigen is a component of a heterogeneous mixture.
Claim 45
Dependent Claim
16 words
The method of claim 41 wherein said heterogeneous mixture is an extract of human melanoma cells.
Claim 46
Dependent Claim
12 words
The method of claim 41 wherein said antigen is human melanoma-associated proteoglycan.
Claim 47
Dependent Claim
22 words
The method of claim 32 wherein said first epitope is an epitope of human melanoma-associated proteoglycan that binds to monoclonal antibody 9.2.27.
Claim 48
Dependent Claim
16 words
The method of claim 32 wherein the antigen is a component of an enriched antigen mixture.
Claim 49
Dependent Claim
18 words
The method of claim 45 wherein the antigen represents at least 1% (w/w) of the enriched antigen mixture.
Claim 50
Dependent Claim
12 words
The method of claim 32 wherein the antigen is a soluble antigen.
Claim 51
Dependent Claim
12 words
The method of claim 32 wherein the antigen is a tumor-associated antigen.
Claim 52
Independent Claim
73 words
A method for producing monoclonal anti-idiotypic antibodies, comprising the steps of: absorbing a first antibody onto insolubilized protein A, thereby forming an immunosorbent; immunizing an animal with said immunosorbent; fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing monoclonal antibodies to an idiotope of said first antibody; culturing said hybridomas to produce said monoclonal antibodies; and collecting said monoclonal antibodies as a product of said hybridomas.
Claim 53
Independent Claim
73 words
A method for producing monoclonal anti-idiotypic antibodies, comprising the steps of: absorbing a first antibody onto insolubilized protein A, thereby forming an immunosorbent; immunizing an animal with said immunosorbent; fusing spleen cells from said immunized animal to myeloma cells to form hybridomas capable of producing monoclonal antibodies to an idiotope of said first antibody; culturing said hybridomas to produce said monoclonal antibodies; and collecting said monoclonal antibodies as a product of said hybridomas.
Claim 54
Dependent Claim
45 words
The method of claim 49, including, after the step of absorbing a first antibody onto insolubilized protein A, thereby forming an immunosorbent, the step of combining a second antibody which is recognized by said first antibody with said immunosorbent, thereby forming an insolubilized immune complex.
Claim 55
Dependent Claim
17 words
The method of claim 50 wherein the first antibody is anti-globulin and the second antibody is anti-antigen.
Claim 56
Dependent Claim
20 words
The method of claim 52 wherein the insolubilized protein A is comprised of protein A bound to an inert matrix.
Claim 57
Dependent Claim
13 words
The method of claim 52 wherein the inert matrix is a polysaccharide matrix.
Claim 58
Dependent Claim
12 words
The method of claim 53 wherein the inert polysaccharide matrix is Sepharose®.
Claim 59
Dependent Claim
15 words
The method of claim 52 wherein the inert matrix is in the form of particles.
Claim 60
Dependent Claim
14 words
The method of claim 55 wherein the particles are in the form of beads.
Claim 61
Dependent Claim
23 words
The method of claim 51, comprising, after the step of absorbing the antibody, the additional step of crosslinking the antibody and protein A.
Claim 62
Dependent Claim
22 words
The method of claim 51, comprising, after the step of combining the antigen, the additional step of crosslinking the antibody and antigen.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1992-11-06 | 1993-05-08 | 1993-11-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-11-06 | 1997-05-08 | 1997-11-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-11-06 | 2001-05-08 | 2001-11-07 | $7700.00 | Window Closed / Lapsed |