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KMID : 0617319950050010090
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Journal of Pharmacetical Sceiences Ewha Womans University
1995 Volume.5 No. 1 p.90 ~ p.99
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Monoclonal Antibodies Reactive With Interleukin-2 Receptor β-Chain Generated by Using a Synthetic Polypeptide as Antigen
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이희구/Lee, Hee Gu
이홍수/이인애/최용경/최인성/정태화/김길현/Lee, Hong-Soo/Lee, In-Ae/Choe, Yong-Kyung/Choe, In-Seong/Chung, Tai-Wha/Kim, Kilhyoun
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Abstract
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Hybridoma producing monoclonal antibodies reactive with the ß-chain of interleukin-2 receptor (IL-2R) were generated from mice immunized with a synthetic pepride of 19 amino acids. The synthetic peptide constitutes strongly hydrophilic region of extracellular domain of the IL-2R molecules(residues 32-50) as revealed by hydrophathy plot analysis. Hybridomas were screened by ELISA using Hut102 cells and the synthetic peptide as antigenic moiety, and by measuring the binding activity of antibodies to the Hut102 cell surface that was monitored by a flow cytometer. The antibodies were further screened by their inhibitory effect against the interaction between IL-2R and interleukin-2(IL-2). Antibodies were further characterized by immunoprecipitation followed by autoradiography to reveal that the antibodies recongnize the 75 KD protein molecules expressed on the Hut 102 cell surface and human peripheral blood mononuclear cells(PBMC) which confirms the antigenic moiety to be ß-chain of IL-2 recepter molecules.
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