Rise and fall of an anti-MUC1 specific antibody.
Rise and fall of an anti-MUC1 specific antibody.
复制标题
抗MUC1特异性抗体的上升和下降。
DOI：
10.1371/journal.pone.0015921
复制
发表时间：
2011-01-14
期刊：
影响因子：
3.7
通讯作者：
Dübel S
中科院分区：
文献类型：
--
作者：
Thie H;Toleikis L;Li J;von Wasielewski R;Bastert G;Schirrmann T;Esteves IT;Behrens CK;Fournes B;Fournier N;de Romeuf C;Hust M;Dübel S
迄今为止，对 MUC1（一种在乳腺癌和卵巢癌上过度表达的跨膜蛋白）具有良好亲和力和特异性的人类抗体非常难以产生，因此是一个有希望的治疗靶点。从免疫文库中分离出人类 scFv 抗体，该免疫文库源自接受 MUC1 免疫的乳腺癌患者。抗 MUC1 scFv 与乳腺癌样本的 228 个组织切片中超过 80% 的肿瘤细胞发生反应，而与大量非肿瘤组织的反应性非常低。通过诱变和噬菌体展示，scFv 的亲和力增加了 500 倍，达到 5,7×10−10 M。血清中的半衰期从 1 天以下提高到 4 周以上，并且与单个 scFv 的二聚化趋势相关。与 T47D 和 MCF-7 哺乳动物癌细胞系结合的 scFv 被重新克隆为 scFv-Fc 和 IgG 格式，导致一种结合物的亲和力降低。使用 MCF-7 和 OVCAR 肿瘤细胞在小鼠异种移植模型中测试了具有最高亲和力的 IgG 变体。然而，实验表明肿瘤生长没有显着下降，存活率也没有显着提高。为了研究异种移植实验失败的原因，使用 MCF-7 和 OVCAR3 靶细胞对 ADCC 进行了体外分析，结果发现，如 MCF-7 细胞检测到的那样，ADCC 较低，可能是由于内化所致。从免疫文库开始，然后进行亲和力成熟的抗体噬菌体展示是一种针对困难靶标产生高亲和力人类抗体的强大策略，在这种情况下，通过创建对由四个氨基酸组成的非常小的表位具有亚纳摩尔亲和力的高度特异性抗体来证明。尽管有这些“同类最佳”的结合参数，但该抗体的治疗成功却受到目标生物学的阻碍。
So far, human antibodies with good affinity and specificity for MUC1, a transmembrane protein overexpressed on breast cancers and ovarian carcinomas, and thus a promising target for therapy, were very difficult to generate. A human scFv antibody was isolated from an immune library derived from breast cancer patients immunised with MUC1. The anti-MUC1 scFv reacted with tumour cells in more than 80% of 228 tissue sections of mamma carcinoma samples, while showing very low reactivity with a large panel of non-tumour tissues. By mutagenesis and phage display, affinity of scFvs was increased up to 500fold to 5,7×10−10 M. Half-life in serum was improved from below 1 day to more than 4 weeks and was correlated with the dimerisation tendency of the individual scFvs. The scFv bound to T47D and MCF-7 mammalian cancer cell lines were recloned into the scFv-Fc and IgG format resulting in decrease of affinity of one binder. The IgG variants with the highest affinity were tested in mouse xenograft models using MCF-7 and OVCAR tumour cells. However, the experiments showed no significant decrease in tumour growth or increase in the survival rates. To study the reasons for the failure of the xenograft experiments, ADCC was analysed in vitro using MCF-7 and OVCAR3 target cells, revealing a low ADCC, possibly due to internalisation, as detected for MCF-7 cells. Antibody phage display starting with immune libraries and followed by affinity maturation is a powerful strategy to generate high affinity human antibodies to difficult targets, in this case shown by the creation of a highly specific antibody with subnanomolar affinity to a very small epitope consisting of four amino acids. Despite these “best in class” binding parameters, the therapeutic success of this antibody was prevented by the target biology.
登录
查看更多内容
影响因子：
14.9
作者：
Ferreira CS;Cheung MC;Missailidis S;Bisland S;Gariépy J
通讯作者：
Gariépy J
影响因子：
5.6
作者：
Abhinandan, K. R.;Martin, Andrew C. R.
通讯作者：
Martin, Andrew C. R.
影响因子：
3.3
作者：
Beatty, Pamela L.;Narayanan, Sowmya;Finn, Olivera J.
通讯作者：
Finn, Olivera J.
影响因子：
--
作者：
Blockzjil, A;Nilsson, K;Nilsson, O
通讯作者：
Nilsson, O
影响因子：
3.5
作者：
Arndt, MAE;Krauss, R;Rybak, SM
通讯作者：
Rybak, SM