Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function.
Identification of antibody glycosylation structures that predict monoclonal antibody Fc-effector function.
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鉴定预测单克隆抗体FC效应功能的抗体糖基化结构。
DOI：
10.1097/qad.0000000000000444
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发表时间：
2014-11-13
期刊：
影响因子：
--
通讯作者：
Alter G
中科院分区：
文献类型：
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作者：
Chung AW;Crispin M;Pritchard L;Robinson H;Gorny MK;Yu X;Bailey-Kellogg C;Ackerman ME;Scanlan C;Zolla-Pazner S;Alter G
目的：确定可预测片段结晶(Fc)介导的抗HIV效应功能的单抗(MAb)特性。来自中国仓鼠卵巢细胞或Epstein-Barr病毒永生化的小鼠异基因骨髓瘤的单抗，具有HIV包膜关键区域的特异性，包括gp120-V2、gp120-V3环、gp120-CD4+结合部位和gp41特异性抗体，以确定抗体的可变和恒定结构域特征在驱动强大的Fc效应功能中的相对贡献。检测抗体与gp140SF162的亲和力、抗体依赖的细胞毒性(ADCC)、抗体依赖的细胞吞噬功能(ADCP)以及与FcgRIIa、FcgRIIb和FcgRIIIa受体的结合能力。用高效液相色谱法测定抗体的葡聚糖谱。单抗的特异性和亲和力都不能决定Fc效应器功能的效力。FcgRIIIa结合强烈预测ADCC，半乳糖含量降低与ADCP呈负相关，而N-羟基神经氨酸结构表现出增强的ADCP。此外，添加了半乳糖的双天线糖链预测与FcgRIIIa和ADCC活性的结合增强，与mAb的特异性无关。我们的研究指出，特定的Fc-葡聚糖结构可以选择性地促进Fc-效应器的功能，而不依赖于抗体的特异性。此外，我们还证明了抗体多糖结构与增强的ADCP活性有关，这是一种新出现的Fc效应功能，可能有助于控制和清除HIV感染。
To determine monoclonal antibody (mAb) features that predict fragment crystalizable (Fc)-mediated effector functions against HIV. Monoclonal antibodies, derived from Chinese hamster ovary cells or Epstein–Barr virus-immortalized mouse heteromyelomas, with specificity to key regions of the HIV envelope including gp120-V2, gp120-V3 loop, gp120-CD4+ binding site, and gp41-specific antibodies, were functionally profiled to determine the relative contribution of the variable and constant domain features of the antibodies in driving robust Fc-effector functions. Each mAb was assayed for antibody-binding affinity to gp140SF162, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP) and for the ability to bind to FcgRIIa, FcgRIIb and FcgRIIIa receptors. Antibody glycan profiles were determined by HPLC. Neither the specificity nor the affinity of the mAbs determined the potency of Fc-effector function. FcgRIIIa binding strongly predicted ADCC and decreased galactose content inversely correlated with ADCP, whereas N-glycolylneuraminic acid-containing structures exhibited enhanced ADCP. Additionally, the bi-antenary glycan arm onto which galactose was added predicted enhanced binding to FcgRIIIa and ADCC activity, independent of the specificity of the mAb. Our studies point to the specific Fc-glycan structures that can selectively promote Fc-effector functions independently of the antibody specificity. Furthermore, we demonstrated antibody glycan structures associated with enhanced ADCP activity, an emerging Fc-effector function that may aid in the control and clearance of HIV infection.