US4775638A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Single vial technique for radiolabeling protein
Assignee: Fujirebio Inc
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Filing: 1985-05-08
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Granted: 1988-10-04
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Expires: 2005-05-08
Abstract
Abstract A method for radiolabeling protein such as antibody which is performed in a single reaction vessel is described. A sealed reaction vessel having a port for addition and withdrawal of reagents preferably by syringe is used. Reagents for coupling radioisotope to the protein are added to the vessel. For radioiodination procedures, vessels can be pre-coated with the iodine coupling agent iodgen. The protein and the radioisotope are then added to the vessel and the radiolabel reaction allowed to proceed. After the reaction is complete, a resin is added to the vessel to adsorb the uncoupled radioisotope. The entire reaction mixture is then withdrawn from the vessel and the resin is separated from the protein preferably by sterile filtration.
Inventors
- 👤 Hidde J. Haisma
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 18:51
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/534; Y; Y10; Y10S; Y10S436/00; Y10S436/804; Y10S436/807; Y10S436/808
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
109 words
A method of labeling an antibody with a radioisotope, comprising the steps of: (a) providing a sealed vessel containing a reagent which catalyst the coupling of the radioisotope to protein, the vessel having means for the introduction and withdrawal of reagents when sealed; (b) introducing into the vessel the radioisotope and a buffer solution of the antibody to be labeled; (c) incubating the resulting reaction mixture under conditions which permit the labeling of the antibody; (d) introducing a resin which adsorbs the radioisotope which is not bound to antibody into the vial; (e) withdrawing the resin and the reaction mixture; and (f) separating the resin and the radiolabeled antibody.
Claim 2
Independent Claim
109 words
A method of labeling an antibody with a radioisotope, comprising the steps of: (a) providing a sealed vessel containing a reagent which catalyst the coupling of the radioisotope to protein, the vessel having means for the introduction and withdrawal of reagents when sealed; (b) introducing into the vessel the radioisotope and a buffer solution of the antibody to be labeled; (c) incubating the resulting reaction mixture under conditions which permit the labeling of the antibody; (d) introducing a resin which adsorbs the radioisotope which is not bound to antibody into the vial; (e) withdrawing the resin and the reaction mixture; and (f) separating the resin and the radiolabeled antibody.
Claim 3
Dependent Claim
12 words
A method of claim 1, wherein the antibody is a monoclonal antibody.
Claim 4
Dependent Claim
16 words
A method of claim 1, wherein the radioisotope is 123 I, 125 I, or 131 I.
Claim 5
Dependent Claim
14 words
A method of claim 1, wherein the radioisotope is 111 In or 67 Ga.
Claim 6
Dependent Claim
32 words
A method of claim 1, wherein the reaction vessel is sealed with a cap penetrable by a needle and reagents are introduced and withdrawn from the vessel by means of a syringe.
Claim 7
Dependent Claim
16 words
A method of claim 1, wherein the resin and the antibody are separated by sterile filtration.
Claim 8
Independent Claim
122 words
A method of iodinating antibody comprising the steps of: (a) placing an oxidizing reagent which catalyzes the coupling of iodine to antibody into a sealable reaction vessel, the vessel having means for introduction and withdrawal of reagents when sealed; (b) sealing the vessel; (c) introducing radioactive iodine and a buffer solution containing the antibody to be iodinated into the vessel to form a reaction mixture; (d) incubating the resulting reaction mixture under conditions which permit iodination of the antibody; (e) introducing into the vessel a resin which absorbs iodine to adsorb iodine which is not bound to the antibody; (f) withdrawing the reaction mixture and the resin from the vessel; and (g) separating the resin and the reaction mixture containing iodinated antibody.
Claim 9
Independent Claim
122 words
A method of iodinating antibody comprising the steps of: (a) placing an oxidizing reagent which catalyzes the coupling of iodine to antibody into a sealable reaction vessel, the vessel having means for introduction and withdrawal of reagents when sealed; (b) sealing the vessel; (c) introducing radioactive iodine and a buffer solution containing the antibody to be iodinated into the vessel to form a reaction mixture; (d) incubating the resulting reaction mixture under conditions which permit iodination of the antibody; (e) introducing into the vessel a resin which absorbs iodine to adsorb iodine which is not bound to the antibody; (f) withdrawing the reaction mixture and the resin from the vessel; and (g) separating the resin and the reaction mixture containing iodinated antibody.
Claim 10
Dependent Claim
12 words
A method of claim 7, wherein the antibody is a monoclonal antibody.
Claim 11
Dependent Claim
25 words
A method of claim 7, wherein the oxidizing reagent which catalyzes the coupling of iodine to antibody is 1,3,4,6-tetrachloro-3,6-diphenylglycuril, chloroamine T, lactoperoxidase or iodine monochloride.
Claim 12
Dependent Claim
20 words
A method of claim 7, wherein the radioactive iodine is I 125 , I 131 , or I 123 .
Claim 13
Dependent Claim
24 words
A method of claim 7, wherein the buffer solution containing the antibody to be iodinated is a 0.1M borate buffer at about ph 8.0-8.5.
Claim 14
Dependent Claim
20 words
A method of claim 7, wherein the resin comprises a styrene divinyl benzene copolymer lattice having attached quarternary ammonium groups.
Claim 15
Dependent Claim
19 words
A method of claim 7, wherein the resin is added to the reaction vessel together with a carrier protein.
Claim 16
Dependent Claim
29 words
A method of claim 7, wherein the resin is separated from the iodinated antibody by filtration through a filter having a pore size less than the resin particle size.
Claim 17
Independent Claim
156 words
A method of iodinating monoclonal antibody, comprising the steps of: (a) providing a vial sealed under a vacuum by a cap penetrable by a needle, the inner surface of the vial being coated with 1,3,4,6-tetrachloro-3,6-diphenylglycuril; (b) injecting into the vial a buffer solution of I 123 , I 125 , or I 131 in sodium hydroxide; (c) injecting into the vial a buffer solution of monoclonal antibody to be iodinated in a borate buffer at ph 8.0-8.5; (d) incubating the resulting reaction mixture under conditions which permit iodination of the antibody; (e) injecting into the vial a mixture comprising a resin which absorbs iodine that is not coupled to the antibody and a carrier protein; (f) withdrawing the reaction mixture and the resin from the vial into a syringe; and (g) separating the resin from the reaction mixtrure containing iodinated antibody by injecting the mixture through a filter having a pore size of about 0.22 microns.
Claim 18
Independent Claim
156 words
A method of iodinating monoclonal antibody, comprising the steps of: (a) providing a vial sealed under a vacuum by a cap penetrable by a needle, the inner surface of the vial being coated with 1,3,4,6-tetrachloro-3,6-diphenylglycuril; (b) injecting into the vial a buffer solution of I 123 , I 125 , or I 131 in sodium hydroxide; (c) injecting into the vial a buffer solution of monoclonal antibody to be iodinated in a borate buffer at ph 8.0-8.5; (d) incubating the resulting reaction mixture under conditions which permit iodination of the antibody; (e) injecting into the vial a mixture comprising a resin which absorbs iodine that is not coupled to the antibody and a carrier protein; (f) withdrawing the reaction mixture and the resin from the vial into a syringe; and (g) separating the resin from the reaction mixtrure containing iodinated antibody by injecting the mixture through a filter having a pore size of about 0.22 microns.
Claim 19
Dependent Claim
25 words
A method of claim 15, wherein the resin is an anion exchange resin comprising a styrene divinyl benzene copolymer lattice having attached quarternary ammonium groups.
Claim 20
Dependent Claim
13 words
A method of claim 15, wherein the carrier protein is bovine serum albumin.
Claim 21
Dependent Claim
55 words
A kit for radioiodinating protein, comprising; a. a vacuum sealed reaction vial the inner surfaces of which is coated with 3,4,6-tetrachloro 3,6-diphenylglycuril, the vial having means for injection and withdrawal of reagents by syringe when the vial is sealed; and b. a vial containing a suspension of a resin which adsorbs iodine in buffer solution.
Claim 22
Dependent Claim
55 words
A kit for radioiodinating protein, comprising; a. a vacuum sealed reaction vial the inner surfaces of which is coated with 3,4,6-tetrachloro 3,6-diphenylglycuril, the vial having means for injection and withdrawal of reagents by syringe when the vial is sealed; and b. a vial containing a suspension of a resin which adsorbs iodine in buffer solution.
Claim 23
Dependent Claim
18 words
A kit of claim 18, further comprising: c. a filter having a pore size of about 0.22 microns.
Claim 24
Dependent Claim
20 words
A kit of claim 18, wherein the resin comprises a styrene divinyl benzene copolymer lattice having attached quarternary ammonium groups.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1991-10-04 | 1992-04-04 | 1992-10-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-10-04 | 1996-04-04 | 1996-10-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-10-04 | 2000-04-04 | 2000-10-04 | $7700.00 | Window Closed / Lapsed |