Generation of inhibitory monoclonal antibodies targeting matrix metalloproteinase-14 by motif grafting and CDR optimization
Generation of inhibitory monoclonal antibodies targeting matrix metalloproteinase-14 by motif grafting and CDR optimization
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DOI：
10.1093/protein/gzw070
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发表时间：
2017-02-01
影响因子：
2.4
通讯作者：
Ge, Xin
中科院分区：
文献类型：
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作者：
Nam, Dong Hyun;Fang, Kuili;Ge, Xin
基质金属蛋白酶-14 （Matrix metalloproteinase-14, MMP-14）在肿瘤转移过程中起重要作用，广谱MMP复合抑制剂在临床试验中的失败提示其选择性至关重要。通过将MMP-14特异性抑制基序嫁接到抗体支架的互补决定区(CDR)-H3中，并对CDR-H3旁边的CDR和序列进行优化，我们分离出了一个结合亲和力为8.3 nM的Fab 1F8，其抑制效能比肽抑制剂提高了1000倍。酵母表面展示和荧光活化细胞分选结果表明，1F8对MMP-14具有高度选择性，并与TIMP-2竞争结合到MMP-14的催化结构域。将低亲和肽抑制剂转化为高效抗体，所述方法可用于开发其他具有治疗意义的抑制性抗体。
Matrix metalloproteinase-14 (MMP-14) plays important roles in cancer metastasis, and the failures of broad-spectrum MMP compound inhibitors in clinical trials suggested selectivity is critical. By grafting an MMP-14 specific inhibition motif into complementarity determining region (CDR)-H3 of antibody scaffolds and optimizing other CDRs and the sequences that flank CDR-H3, we isolated a Fab 1F8 showing a binding affinity of 8.3 nM with >1000-fold enhancement on inhibition potency compared to the peptide inhibitor. Yeast surface display and fluorescence-activated cell sorting results indicated that 1F8 was highly selective to MMP-14 and competed with TIMP-2 on binding to the catalytic domain of MMP-14. Converting a low-affinity peptide inhibitor into a high potency antibody, the described methods can be used to develop other inhibitory antibodies of therapeutic significance.