通过基序嫁接和互补决定区优化产生靶向基质金属蛋白酶-14的抑制性单克隆抗体。
Generation of inhibitory monoclonal antibodies targeting matrix metalloproteinase-14 by motif grafting and CDR optimization.
作者信息
Nam Dong Hyun, Fang Kuili, Rodriguez Carlos, Lopez Tyler, Ge Xin
机构信息
Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA.
Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA
出版信息
Protein Eng Des Sel. 2017 Feb;30(2):113-118. doi: 10.1093/protein/gzw070. Epub 2016 Dec 15.
Abstract
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.
摘要
基质金属蛋白酶-14（MMP-14）在癌症转移中起重要作用，而广谱MMP复合抑制剂在临床试验中的失败表明选择性至关重要。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，并优化其他CDR和CDR-H3侧翼序列，我们分离出一种Fab 1F8，其结合亲和力为8.3 nM，与肽抑制剂相比，抑制效力提高了1000倍以上。酵母表面展示和荧光激活细胞分选结果表明，1F8对MMP-14具有高度选择性，并在与MMP-14催化结构域结合时与TIMP-2竞争。将低亲和力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。
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