US4742159A
Expired / Lapsed
US
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Digitalis antibodies, process for the preparation thereof and the use thereof for the therapy of digitalis intoxications
Assignee: Roche Diagnostics GmbH
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Filing: 1983-12-19
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Granted: 1988-05-03
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Expires: 2003-12-19
Abstract
Abstract The present invention provides a process for the preparation of digitalis antibodies in which appropriate mammals are immunized with a digoxin bound to protein, the animal serum is obtained, the immune globulin-containing protein fraction is separated in known manner, the immunologically-active globulins are adsorbed on an immunologically-active column and separated from the other proteins, the antibodies are again eluted from the column and the Fab fractions are split with papain and purified, wherein, as immunologically-active adsorbent, there is used an inorganic matrix of large surface area to which digitoxin aldehyde is bound via a spacer which cannot be split with papain and the splitting off of the Fab fraction from the antibodies is carried out on the matrix. The present invention also provides digitalis antibodies obtained by this process, which antibodies can be used for the therapy of digitalis intoxications and for preparing immunological reagents for the determination of digitalis glycosides.
Inventors
- 👤 Hans-Georg Batz
- 👤 Herbert Jungfer
- 👤 Helmut Lenz
- 👤 Albert Roder
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 13:19
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/16; A; A61; A61P; A61P39/00; A61P39/02; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/94; G01N33/9453; A61K; A61K38/00; Y; Y10; Y10S; Y10S424/00; Y10S424/809; Y10S530/00; Y10S530/866
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
162 words
In a process for the preparation of Fab fragments of digitalis antibodies in which appropriate mammals are immunised with an agent for immunizing comprising a digoxin bound to protein, the animal serum obtained, the immune globulin-containing protein fraction is separated in known manner, the immunologically-active globulins adsorbed on an immunologically-active adsorbent in a column and separated from the other proteins, the antibodies again eluted from the column and the Fab fractions split with papain and purified wherein the improvement comprises absorbing the digitalis antibodies on an immunologically-active absorbent which comprises an inorganic matrix to which digitoxin aldehyde is bound via a spacer which cannot be split with papain, treating the matrix with papain so as to split the absorbed antibodies into Fab and Fc fragments, washing the column so as to remove the Fc fragments, washing the column so as to remove the Fc fragments and excess papain, and desorbing the Fab fragments bound to the matrix with a suitable eluting agent.
Claim 2
Dependent Claim
162 words
In a process for the preparation of Fab fragments of digitalis antibodies in which appropriate mammals are immunised with an agent for immunizing comprising a digoxin bound to protein, the animal serum obtained, the immune globulin-containing protein fraction is separated in known manner, the immunologically-active globulins adsorbed on an immunologically-active adsorbent in a column and separated from the other proteins, the antibodies again eluted from the column and the Fab fractions split with papain and purified wherein the improvement comprises absorbing the digitalis antibodies on an immunologically-active absorbent which comprises an inorganic matrix to which digitoxin aldehyde is bound via a spacer which cannot be split with papain, treating the matrix with papain so as to split the absorbed antibodies into Fab and Fc fragments, washing the column so as to remove the Fc fragments, washing the column so as to remove the Fc fragments and excess papain, and desorbing the Fab fragments bound to the matrix with a suitable eluting agent.
Claim 3
Dependent Claim
15 words
Process according to claim 1, wherein porous glass is used as the inorganic matrix material.
Claim 4
Dependent Claim
12 words
Process according to claim 1, wherein triethoxysilylpropylamine is used as the spacer.
Claim 5
Dependent Claim
33 words
Process according to claim 1, wherein digitoxin aldehyde, obtained by the oxidation of digitoxin with periodate, is bound to the matrix as Schiffs base and the Schiffs base is reduced with sodium borohydride.
Claim 6
Dependent Claim
18 words
Process according to claim 1, wherein digoxin-glutaryl-O-hydroxysuccinimide bound to albumin is used as the agent for the immunizing.
Claim 7
Dependent Claim
21 words
Process according to claim 1, wherein the antibody containing globulin fraction is obtained by ammonium sulfate precipitation from the animal serum.
Claim 8
Dependent Claim
33 words
Process according to claim 1, wherein the aborption of the antibodies and the the removal of the Fc fragments and excess papain is effected with a phosphate/sodium chloride solution buffered to pH 7.
Claim 9
Dependent Claim
23 words
Process according to claim 1, wherein the Fab fragments desorbed from the column are further purified by chromatography on gels and/or ion exchangers.
Claim 10
Dependent Claim
23 words
Process according to claim 1 wherein the matrix is porous glass and the digitoxin aldehyde is bound thereto by triethoxysilylpropylamine as the spacer.
References Cited (Backward Citations)
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