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抗蓖麻毒素单克隆抗体:对毒素功能的影响。

Monoclonal antibodies against ricin: effects on toxin function.

作者信息

Colombatti M, Pezzini A, Colombatti A

出版信息

Hybridoma. 1986 Spring;5(1):9-19. doi: 10.1089/hyb.1986.5.9.

Abstract

Monoclonal antibodies against ricin toxin were produced and some of their properties investigated. Antibodies 196 C12 and 197 C7 raised against A-chain reacted with a CnBr fragment probably comprised between amino acid 254 and 262. Antibodies 193 A9, 196 A3, and 191 B7 recognized a 6-7 kD CnBr peptide. A second set of antibodies was raised against whole inactivated ricin. Most of them bound in a solid phase radioimmunobinding assay only to ricin and few had a low activity against purified A-chain. Different effects were noted on toxin action in cultured leukemic cells. If cells were preincubated with ricin followed by antibodies, MAb 207 E5 and 216 B3 had a strong enhancing effect on toxin action. If antibodies and toxin were mixed and then added to sensitive cells, antibody 207 E5 gave a strong protection while 216 B3 maintained its enhancing activity. The effect of antibody 216 B3 was further investigated by quantitative cloning experiments which showed that toxin had a fivefold enhancement in its activity by a preincubation with this antibody. Binding of fluoresceinated ricin to leukemic target cells was inhibited by a preincubation with antibody 207 E5 while antibody 216 B3 had no effect.

摘要

制备了抗蓖麻毒素的单克隆抗体,并对其一些特性进行了研究。针对A链产生的抗体196 C12和197 C7与一个可能包含氨基酸254至262之间的溴化氰片段发生反应。抗体193 A9、196 A3和191 B7识别出一个6 - 7 kD的溴化氰肽段。制备了第二组针对完全灭活蓖麻毒素的抗体。在固相放射免疫结合试验中,它们大多数仅与蓖麻毒素结合,少数对纯化的A链有低活性。在培养的白血病细胞中观察到对毒素作用的不同影响。如果细胞先用蓖麻毒素预孵育,然后加入抗体,单克隆抗体207 E5和216 B3对毒素作用有强烈的增强作用。如果将抗体和毒素混合,然后加入敏感细胞,抗体207 E5给予强烈的保护作用,而216 B3保持其增强活性。通过定量克隆实验进一步研究了抗体216 B3的作用,结果表明毒素与该抗体预孵育后其活性增强了五倍。用抗体207 E5预孵育可抑制荧光素化蓖麻毒素与白血病靶细胞的结合,而抗体216 B3则无此作用。

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