通过噬菌体展示分离的重组人抗Rh D Fab片段的序列和特异性分析
Sequence and specificity analysis of recombinant human Fab anti-Rh D isolated by phage display.
作者信息
Miescher S, Vogel M, Biaggi C, Ramseyer V, Hustinx H, Eicher N, Imboden M A, Spycher M O, Amstutz H, Stadler B M
机构信息
ZLB Central Laboratory, Blood Transfusion Service SRC, Inselspital, Bern, Switzerland.
出版信息
Vox Sang. 1998;75(4):278-87.
BACKGROUND AND OBJECTIVES
Hyperimmune anti-Rh D serum is worldwide in short supply. As a first step to develop an alternative source of Rh D antibodies, we describe in this study the isolation and characterization of recombinant anti-Rh D Fab fragments.
MATERIALS AND METHODS
Peripheral blood mononuclear cells harvested from a hyperimmunized donor were used to construct two combinatorial Fab libraries. Phages expressing these Fab fragments were selected on whole red blood cells followed by testing of positive clones in an indirect hemagglutination assay.
RESULTS
Individual Fab clones are of high affinity and competitively inhibit the binding of a registered anti-D immunoglobulin. The Fab clones are also specific against the partial D phenotypes, Rh33, DIII, DIVa, DIVb, DVa, and DVII. The 13 different but highly homologous clones express preferentially VH3 segments.
CONCLUSION
These Fab fragments show potential for the development of a new generation of therapeutic anti-Rh D reagents.
背景与目的
超免疫抗Rh D血清在全球范围内供应短缺。作为开发Rh D抗体替代来源的第一步，我们在本研究中描述了重组抗Rh D Fab片段的分离与特性鉴定。
材料与方法
从一名超免疫供体采集的外周血单个核细胞用于构建两个组合Fab文库。在全红细胞上选择表达这些Fab片段的噬菌体，随后在间接血凝试验中检测阳性克隆。
结果
单个Fab克隆具有高亲和力，并竞争性抑制已注册抗-D免疫球蛋白的结合。Fab克隆对部分D表型、Rh33、DIII、DIVa、DIVb、DVa和DVII也具有特异性。13个不同但高度同源的克隆优先表达VH3片段。
结论
这些Fab片段显示出开发新一代治疗性抗Rh D试剂的潜力。
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