Novel conformation‐selective monoclonal antibodies against apoA‐I amyloid fibrils
Novel conformation‐selective monoclonal antibodies against apoA‐I amyloid fibrils
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针对 apoA-I 淀粉样原纤维的新型构象选择性单克隆抗体
DOI：
10.1111/febs.15487
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发表时间：
2021
期刊：
影响因子：
5.4
通讯作者：
Hiroyuki Saito
中科院分区：
文献类型：
--
作者：
Takashi Ohgita;Yuki Furutani;Miyu Nakano;Megumi Hattori;Ayane Suzuki;Miho Nakagawa;Sera Naniwa;Izumi Morita;Hiroyuki Oyama;Kazuchika Nishitsuji;Norihiro Kobayashi;Hiroyuki Saito
人载脂蛋白A-I（apoA-I）（血浆高密度脂蛋白的主要蛋白）中的爱荷华州（G26 R）突变与系统性淀粉样变性相关，携带该突变的apoA-I N末端1-83片段具有形成淀粉样纤维的强烈倾向。在这里，我们产生并表征了对apoA-I淀粉样纤维显示选择性反应性的新型单克隆抗体（mAb）。通过用apoA-I 1 β 83/G26 R原纤维与血蓝蛋白缀合免疫BALB/c和A/J小鼠并产生杂交瘤，获得了4种IgM类mAb。产生的mAb对由apoA-I的1-83片段形成的淀粉样纤维表现出强反应性，但对单体1-83片段或全长apoA-I不表现出强反应性。mAb与apoA-I原纤维的表观解离常数确定在nM范围内。时间依赖性聚集测定表明，mAb优先与成熟原纤维反应，而不是与apoA-I 1-83/G26 R形成的非原纤维聚集体反应。此外，使用apoA-I 1 β 83/G26 R的缺失或脯氨酸取代变体的斑点印迹和ELISA表明，所产生的mAb与apoA-I淀粉样蛋白原纤维中的共同结构特征反应。事实上，mAb还识别由与apoA-I没有序列同一性的α-突触核蛋白形成的淀粉样纤维。因此，我们新产生的抗apoA-I原纤维单克隆抗体不仅可用于诊断apoA-I相关淀粉样变性，还可作为新型构象选择性抗体用于淀粉样原纤维的结构分析。
The Iowa (G26R) mutation in human apolipoprotein A‐I (apoA‐I), the major protein of plasma high‐density lipoprotein, is associated with systemic amyloidosis, and the N‐terminal 1–83 fragment of apoA‐I carrying this mutation has a strong propensity to form amyloid fibrils. Here, we generated and characterized novel monoclonal antibodies (mAbs) that display selective reactivity to apoA‐I amyloid fibrils. By immunizing BALB/c and A/J mice with apoA‐I 1‒83/G26R fibrils conjugated with hemocyanin and the hybridoma production, four IgM class mAbs were obtained. The generated mAbs exhibited strong reactivity to amyloid fibrils formed by the 1–83 fragment of apoA‐I, but not to the monomeric 1–83 fragment or full‐length apoA‐I. The apparent dissociation constant of the mAbs to apoA‐I fibrils was determined to be within the nM range. A time‐dependent aggregation assay demonstrated that the mAbs preferentially react with mature fibrils over non‐fibrillar aggregates formed by apoA‐I 1–83/G26R. In addition, dot blotting and ELISA using deletion or proline substituted variants of apoA‐I 1‒83/G26R suggest that the generated mAbs react to common structural features in apoA‐I amyloid fibrils. Indeed, the mAbs also recognized amyloid fibrils formed by α‐synuclein that has no sequence identity to apoA‐I. Thus, our newly generated anti‐apoA‐I fibril mAbs may be utilized for not only diagnosis of apoA‐I‐related amyloidosis but also structural analysis of amyloid fibrils as novel conformation‐selective antibodies.
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人类载脂蛋白的淀粉样蛋白形成特性：序列分析和结构见解。
影响因子：
--
作者：
Das M;Gursky O
通讯作者：
Gursky O
DOI：
10.1073/pnas.85.23.8998
发表时间：
1988-12-01
影响因子：
11.1
作者：
FROHMAN, MA;DUSH, MK;MARTIN, GR
通讯作者：
MARTIN, GR
影响因子：
6
作者：
Rowczenio, Dorota;Dogan, Ahmet;Gillmore, Julian D.
通讯作者：
Gillmore, Julian D.
影响因子：
1.9
作者：
Tougaard, Birgitte G.;Pedersen, Katja Venborg;Birn, Henrik
通讯作者：
Birn, Henrik
DOI：
10.1016/j.bbagen.2016.05.040
发表时间：
2016-11
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Ma B;Zhao J;Nussinov R
通讯作者：
Nussinov R