Synthesis and characterization of site-specific porphyrin-antibody conjugates.
Item
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Title
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Synthesis and characterization of site-specific porphyrin-antibody conjugates.
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Identifier
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AAI9417442
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identifier
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9417442
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Creator
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Cai, Shangde.
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Contributor
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Adviser: David K. Lavallee
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Date
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1994
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Language
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English
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Publisher
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City University of New York.
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Subject
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Chemistry, Organic | Chemistry, Inorganic | Chemistry, Biochemistry
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Abstract
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The porphyrin-antibody conjugation strategy reported in this work has potential for use of nuclear medicine due to the robust chelation of radionuclides by porphyrins and the inherent specificity of the antibody-antigen interaction.;A method for regiospecific covalent modification of antibodies at the oligosaccharide moieties of the hinge region of the antibodies was developed and the characterization of the covalent porphyrin-antibody conjugates by using sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) is described.;An asymmetrical bifunctional porphyrin, 5-(4-aminophenyl)-10,15,20,-tris(4-sulfonatophenyl) porphine (H{dollar}\sb{lcub}2{rcub}{dollar}AS{dollar}\sb{lcub}3{rcub}{dollar}P) and its N-p-nitro-benzylated derivative (N-bzHAS{dollar}\sb{lcub}3{rcub}{dollar}P) were synthesized. The former had been used for testing the possibility of attachment of the aniline group of H{dollar}\sb{lcub}2{rcub}{dollar}AS{dollar}\sb{lcub}3{rcub}{dollar}P to the aldehyde group produced by mild oxidation of oligosaccharide moieties in antibodies. The conjugation was verified by SDS-PAGE slab gel which revealed coloration due to the porphyrin at the same position as the antibody's heavy chain band, demonstrating that the porphyrin covalently binds to the heavy chains of antibodies.;To be useful in rapid metallation of the porphyrin-antibody conjugates, the poyphyrin attached to antibodies must be N-substituted. To synthesize an N-substituted porphyrin, it was necessary to protect the aniline functionality during the N-alkylation reaction. The phthalimide group proved stable enough to resist the high temperature required for alkylation of a pyrroleninic nitrogen atom of the poyphyrin. The phthalimide moiety was then deprotected to recover the amino group which is necessary for antibody conjugation.
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Type
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dissertation
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Source
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PQT Legacy CUNY.xlsx
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degree
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Ph.D.
