Patent US4276206A successfully refreshed from Google Patents.
US4276206A
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Immunochemical conjugates: method and composition
Assignee: Scripps Research Institute
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Filing: 1980-02-11
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Granted: 1981-06-30
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Expires: 2000-02-11
Abstract
Abstract Techniques for preparing stable conjugates of protein-D-GL which can then be isolated in pure form are disclosed, using hen egg ovalbumin (OVA) as a prototype protein, and conjugation and purification methods are described which involve (1) application of a coupling method employing m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS) as the coupling reagent, and (2) introduction of biotin moieties into D-GL molecules to allow application of avidinbiotin system for affinity chromatographic purification of conjugates. These methods are applicable to the preparation of D-GL conjugates of insulin and Ragweed Antigen E and are of general applicability.
Inventors
- 👤 David H. Katz
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 22:15
CPC Classifications
C; C07; C07K; C07K1/00; C07K1/10; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/62; A61K47/64; A61K47/645; C07K14/00; C07K14/435; C07K14/575; C07K14/62; A61K39/00; A61K2039/60; A61K2039/6093; A61K38/00; Y; Y10; Y10S; Y10S424/00; Y10S424/805; Y10S424/81; Y10S530/00; Y10S530/806
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
99 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with antigen comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) reacting antigen with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-Antigen or with S-acetylmercaptosuccinic anhydride to form SH-Antigen; (d) reacting MB-D-GL(biotin) with SH-Antigen or reacting SH-D-GL(biotin) with MB-Antigen to form conjugates of D-glutamic acid-D-lysine(biotin) with antigen; and (e) separating said D-glutamic acid-D-lysine(biotin)-antigen conjugate from non-conjugated antigen by affinity chromatography on an avidin-Sepharose chromatographic column.
Claim 2
Independent Claim
99 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with antigen comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) reacting antigen with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-Antigen or with S-acetylmercaptosuccinic anhydride to form SH-Antigen; (d) reacting MB-D-GL(biotin) with SH-Antigen or reacting SH-D-GL(biotin) with MB-Antigen to form conjugates of D-glutamic acid-D-lysine(biotin) with antigen; and (e) separating said D-glutamic acid-D-lysine(biotin)-antigen conjugate from non-conjugated antigen by affinity chromatography on an avidin-Sepharose chromatographic column.
Claim 3
Independent Claim
99 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with insulin comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) recting insulin with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-Insulin or with S-acetylmercaptosuccinic anhydride to form SH-Insulin; (d) reacting MB-D-GL(biotin) with SH-Insulin or reacting SH-D-GL(biotin) with MB-Insulin to form conjugates of D-glutamic acid-D-lysine(biotin) with insulin; and (e) separating said D-glutamic acid-D-lysine(biotin)-insulin conjugate from non-conjugated insulin by affinity chromatography on an avidin-Sepharose chromatographic column.
Claim 4
Independent Claim
99 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with insulin comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) recting insulin with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-Insulin or with S-acetylmercaptosuccinic anhydride to form SH-Insulin; (d) reacting MB-D-GL(biotin) with SH-Insulin or reacting SH-D-GL(biotin) with MB-Insulin to form conjugates of D-glutamic acid-D-lysine(biotin) with insulin; and (e) separating said D-glutamic acid-D-lysine(biotin)-insulin conjugate from non-conjugated insulin by affinity chromatography on an avidin-Sepharose chromatographic column.
Claim 5
Independent Claim
117 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with ragweed antigen E comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) reacting ragweed antigen E with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-ragweed antigen E or with S-acetylmercaptosuccinic anhydride to form SH-ragweed antigen E; (d) reacting MB-D-GL(biotin) with SH-ragweed antigen E or reacting SH-D-GL(biotin) with MB-ragweed antigen E to form conjugates of D-glutamic acid-D-lysine(biotin) with ragweed antigen E; and (e) separating said D-glutamic acid-D-lysine(biotin)-ragweed antigen E conjugate from non-conjugated ragweed antigen E by affinity chromatography on an avidin-Sepharose chromatographic column.
Claim 6
Independent Claim
117 words
The method of preparing therapeutic immunosuppressive conjugates of D-glutamic acid-D-lysine copolymer with ragweed antigen E comprising the steps of: (a) introducing biotin into the D-glutamic acid-D-lysine copolymer molecule to form D-GL(biotin); (b) reacting D-GL(biotin) with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-D-GL(biotin) or with S-acetylmercaptosuccinic anhydride to form SH-D-GL(biotin); (c) reacting ragweed antigen E with either m-maleimidobenzoyl-N-hydroxysuccinimide ester to form MB-ragweed antigen E or with S-acetylmercaptosuccinic anhydride to form SH-ragweed antigen E; (d) reacting MB-D-GL(biotin) with SH-ragweed antigen E or reacting SH-D-GL(biotin) with MB-ragweed antigen E to form conjugates of D-glutamic acid-D-lysine(biotin) with ragweed antigen E; and (e) separating said D-glutamic acid-D-lysine(biotin)-ragweed antigen E conjugate from non-conjugated ragweed antigen E by affinity chromatography on an avidin-Sepharose chromatographic column.
References Cited (Backward Citations)
US3794630A
Cited
US4191668A
Cited
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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) | 1984-06-29 | 1984-12-29 | 1985-06-30 | $2000.00 | Window Closed / Lapsed |
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