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Development of anti-matrix metalloproteinase-2 (MMP-2) nanobodies as potential therapeutic and diagnostic tools.
Nanomedicine: Nanotechnology, Biology and Medicine ( IF 4.2 ) Pub Date : 2019-10-24 , DOI: 10.1016/j.nano.2019.102103
Alessandro Marturano 1 , Maarten L V Hendrickx 2 , Emanuela Falcinelli 1 , Manuela Sebastiano 1 , Giuseppe Guglielmini 1 , Gholamreza Hassanzadeh-Ghassabeh 3 , Serge Muyldermans 4 , Paul J Declerck 2 , Paolo Gresele 1
Affiliation  

Matrix metalloproteinase-2 (MMP-2) is an endopeptidase involved in cardiovascular disease and cancer. To date, no highly selective MMP-2 inhibitors have been identified for potential use in humans. Aim of our work was to apply the nanobody technology to the generation of highly selective inhibitors of human MMP-2 and to assess their effects on platelet function and their applicability as conjugated nanobodies. We constructed a nanobody library after immunising an alpaca with human active MMP-2 and identified, after phage display and screening, one MMP-2 inhibitory nanobody (VHH-29), able to hinder the effects of MMP-2 on platelet activation, and one nanobody not inhibiting MMP-2 activity (VHH-136) which, chemically conjugated to a fluorescent probe, allowed the detection of human MMP-2 by flow-cytometry and immune-cytochemistry. In conclusion, we have generated and characterized two new nanotechnological molecular tools for human MMP-2 which represent promising agents for the study of MMP-2 in cardiovascular pathophysiology.

中文翻译:

抗基质金属蛋白酶2(MMP-2)纳米抗体的开发作为潜在的治疗和诊断工具。

基质金属蛋白酶2(MMP-2)是一种内肽酶,参与心血管疾病和癌症。迄今为止,还没有发现高选择性的MMP-2抑制剂可用于人类。我们的工作目的是将纳米抗体技术应用于人类MMP-2的高选择性抑制剂的产生,并评估其对血小板功能的影响及其作为偶联纳米抗体的适用性。我们用人类活性MMP-2免疫羊驼后,构建了一个纳米抗体文库,并在噬菌体展示和筛选后鉴定了一种MMP-2抑制性纳米抗体(VHH-29),它能够阻止MMP-2对血小板活化的影响,并且一种不抑制MMP-2活性的纳米抗体(VHH-136),通过化学偶联到荧光探针上,可以通过流式细胞仪和免疫细胞化学法检测人MMP-2。综上所述,
更新日期:2019-10-25
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