CONSTRUCTION OF BACTERIOPHAGE EXPRESSING MOUSE MONOCLONAL FAB FRAGMENTS DIRECTED AGAINST THE HUMAN MN-GLYCOPHORIN BLOOD-GROUP ANTIGENS
CONSTRUCTION OF BACTERIOPHAGE EXPRESSING MOUSE MONOCLONAL FAB FRAGMENTS DIRECTED AGAINST THE HUMAN MN-GLYCOPHORIN BLOOD-GROUP ANTIGENS
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DOI：
10.1046/j.1537-2995.1995.35295125736.x
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发表时间：
1995-02-01
期刊：
影响因子：
2.9
通讯作者：
SPITALNIK, SL
中科院分区：
文献类型：
--
作者：
CZERWINSKI, M;SIEGEL, DL;SPITALNIK, SL
背景：MN人血型抗原是血型糖蛋白A氨基端的复合糖肽抗原。已经产生并表征了针对这些抗原的许多不同的小鼠单克隆抗体。构建在噬菌体表面展示抗体Fab片段的组合免疫球蛋白库（Fab噬菌体）代表了开发单克隆试剂、探索对特定抗原的免疫反应多样性以及了解抗体相互作用的分子基础的新方法。与其抗原。研究设计和方法：从三个抗-N（AH 7，N61和N92）和两个抗-M（425/2B和M2 A1）小鼠杂交瘤构建Fab噬菌体。结果：Fab噬菌体抗体与亲本杂交瘤抗体具有相似的免疫学特性。特别是，它们依赖于缓冲液的pH值和唾液酸化的目标antigen.Conclusion：这些结果表明，Fab噬菌体可以提供新的试剂与应用在免疫血液学，并可能是有用的研究人类血型抗原的免疫反应。
Background: The MN human blood group antigens are complex glycopeptide antigens at the amino terminus of glycophorin A. Many different mouse monoclonal antibodies to these antigens have been produced and characterized. The construction of combinatorial immunoglobulin libraries displaying antibody Fab fragments on the surface of bacteriophage (Fab-phage) represents a novel approach for developing monoclonal reagents, for exploring the diversity of the immune response to specific antigens, and for understanding the molecular basis of the interaction of an antibody with its antigen. However, it Is necessary to determine whether Fab fragments displayed on bacteriophage surfaces retain immunologic characteristics similar to the intact antibodies.Study Design and Methods: Fab-phage were constructed from three anti-N (AH7, N61, and N92) and two anti-M (425/2B and M2A1) murine hybridomas. The Fab-phage and parental hybridomas were compared by enzyme-linked immunosorbent assay, Western blotting, and flow cytometry.Results: In each case, the Fab-phage and its parental hybridoma antibody had similar immunologic characteristics. In particular, their dependence on the pH of the buffer and on sialylation of the target antigen was similar.Conclusion: These results suggest that Fab-phage may provide novel reagents with applications in immunohematology and may be useful in the study of the immune response to human blood group antigens.