Selective antibody activation through protease-activated pro-antibodies that mask binding sites with inhibitory domains

I. Ju Chen, Chih Hung Chuang, Yuan Chin Hsieh, Yun Chi Lu, Wen Wei Lin, Chien Chiao Huang, Ta Chun Cheng, Yi An Cheng, Kai Wen Cheng, Yeng Tseng Wang, Fang Ming Chen, Tian Lu Cheng*, Shey-Cherng Tzou

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

29 Scopus citations


Systemic injection of therapeutic antibodies may cause serious adverse effects due to on-target toxicity to the antigens expressed in normal tissues. To improve the targeting selectivity to the region of disease sites, we developed protease-activated pro-antibodies by masking the binding sites of antibodies with inhibitory domains that can be removed by proteases that are highly expressed at the disease sites. The latency-associated peptide (LAP), C2b or CBa of complement factor 2/B were linked, through a substrate peptide of matrix metalloproteinase-2 (MMP-2), to an anti-epidermal growth factor receptor (EGFR) antibody and an anti-tumor necrosis factor-α (TNF-α) antibody. Results showed that all the inhibitory domains could be removed by MMP-2 to restore the binding activities of the antibodies. LAP substantially reduced (53.8%) the binding activity of the anti-EGFR antibody on EGFR-expressing cells, whereas C2b and CBa were ineffective (21% and 9.3% reduction, respectively). Similarly, LAP also blocked 53.9% of the binding activity of the anti-TNF-α antibody. Finally, molecular dynamic simulation showed that the masking efficiency of LAP, C2b and CBa was 33.7%, 10.3% and-5.4%, respectively, over the binding sites of the antibodies. This strategy may aid in designing new protease-activated pro-antibodies that attain high therapeutic potency yet reduced systemic on-target toxicity.

Original languageEnglish
Article number11587
Pages (from-to)1-12
Number of pages12
JournalScientific reports
Issue number1
StatePublished - 14 Sep 2017


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