INSERM, UMR_S 665, 75015 Paris, France.
Cell Mol Life Sci. 2010 Oct;67(19):3371-87. doi: 10.1007/s00018-010-0387-6. Epub 2010 May 11.
Fy blood group antigens are carried by the Duffy antigen receptor for chemokines (DARC), a red cells receptor for Plasmodium vivax broadly implicated in human health and diseases. Recombinant VHHs, or nanobodies, the smallest intact antigen binding fragment derivative from the heavy chain-only antibodies present in camelids, were prepared from a dromedary immunized against DARC N-terminal extracellular domain and selected for DARC binding. A described VHH, CA52, does recognize native DARC on cells. It inhibits P. vivax invasion of erythrocytes and displaces interleukin-8 bound to DARC. The targeted epitope overlaps the well-defined DARC Fy6 epitope. K (D) of CA52-DARC equilibrium is sub-nanomolar, hence ideal to develop diagnostic or therapeutic compounds. Immunocapture by immobilized CA52 yielded highly purified DARC from engineered K562 cells. This first report on a VHH with specificity for a red blood cell protein exemplifies VHHs' potentialities to target, to purify, and to modulate the function of cellular markers.
Fy 血型抗原由趋化因子的 Duffy 抗原受体(DARC)携带,DARC 是红细胞对间日疟原虫的受体,广泛涉及人类健康和疾病。从骆驼科只含有重链的抗体中提取的最小完整抗原结合片段衍生物——重组 VHH 或纳米抗体,是针对 DARC N 端细胞外结构域免疫的骆驼制备的,并针对 DARC 结合进行了选择。一种已描述的 VHH,CA52,确实可以识别细胞上的天然 DARC。它可以抑制间日疟原虫入侵红细胞并取代与 DARC 结合的白细胞介素-8。靶向表位与已明确的 DARC Fy6 表位重叠。CA52-DARC 平衡的 K(D)为亚纳摩尔级,因此非常适合开发诊断或治疗化合物。通过固定化 CA52 的免疫捕获,从工程化的 K562 细胞中获得了高度纯化的 DARC。这是第一篇关于针对红细胞蛋白的 VHH 的报告,展示了 VHH 针对、纯化和调节细胞标志物功能的潜力。