特异于MT1-MMP的亲和肽的筛选，鉴定和表征及其在肿瘤成像中的应用。,Bioconjugate Chemistry

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特异于MT1-MMP的亲和肽的筛选，鉴定和表征及其在肿瘤成像中的应用。
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2019-04-22 , DOI: 10.1021/acs.bioconjchem.9b00220
Xiang Li ₁ , Zheng Ma ₂ , Haoran Wang ₁ , Li Ren ₃ , Dianwen Zhang ₄ , Weiguo Liang ₅ , Guangji Zhang ₁ , Jinrui Zhang ₁ , Dahai Yu ₁ , Xuexun Fang ₁

Affiliation

膜1型基质金属蛋白酶（MT1-MMP）在许多生理和病理过程中，尤其是在肿瘤的侵袭和转移中起着至关重要的作用。该关键分子的生物成像可以在各种应用中找到广泛的用途。MT环是MT1-MMP的独特序列，位于蛋白质表面。在我们以前的研究中，AF7p是一种针对MT1-MMP的MT环结构域的亲和肽，是通过筛选噬菌体展示（Ph.D.）肽库来鉴定的。但是，AF7p的靶标是缺少MT环区域天然构象的合成序列。因此，现实中的结合亲和力和特异性可能不是最佳的。在这项研究中，我们考虑了MT1-MMP分子中MT环区域的3维（3-D）构象，并设计了一种筛选Ph.D的新策略。肽库。通过酶联免疫吸附测定（ELISA）和生物层干涉法（BLI）观察到，我们获得的肽与AF7p相比对WT-MT1-MMP具有更好的结合亲和力。新的肽标记并附着了MT1-MMP表达细胞系HT1080，对细胞没有任何毒性。此外，对于体内成像，MT1-MMP表达较高的荷HT1080荷瘤小鼠在肿瘤部位比MT1-MMP低表达细胞系A549的小鼠积累了更多的Cy5.5-HS7，并且HS7的半衰期更长主要器官的离体成像证实，该抗体比AF7p的抗体要好。这些结果表明使用减法生物淘选策略筛选靶向MT环区域的亲和肽的可行性，而HS7是用于在MT1-MMP阳性肿瘤模型中无创成像MT1-MMP表达的优良探针。

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Screening, Identification, and Characterization of an Affinity Peptide Specific to MT1-MMP and Its Application in Tumor Imaging.

Membrane type-1 matrix metalloproteinase (MT1-MMP) plays a crucial role in many physiological and pathological processes, especially in tumor invasion and metastasis. Bioimaging of this key molecule may find wide usage in various applications. MT-loop is a unique sequence of MT1-MMP and locates in the surface of the protein. In our previous studies, AF7p, an affinity peptide that targeting the MT-loop domain of MT1-MMP, was identified by screening a phage display (Ph.D.) peptide library. However, the target of AF7p is a synthetic sequence which lacked native conformation of the MT-loop region; thus, the binding affinity and specificity in reality may not be optimal. In this study, we considered the 3-dimensional (3-D) conformation of the MT-loop area in the MT1-MMP molecule and designed a novel strategy to screen the Ph.D. peptide library. The peptide we obtained showed a better binding affinity to WT-MT1-MMP than AF7p as observed through enzyme-linked immunosorbent assay (ELISA) and biolayer interferometry (BLI). The new peptide labeled and attached MT1-MMP expression cell lines HT1080 and did not show any toxicity to cells. Furthermore, for in vivo imaging, HT1080 tumor-bearing mice with higher MT1-MMP expression accumulated more Cy5.5-HS7 than mice with MT1-MMP low-expression cell lines A549 at tumor sites, and the half-life of HS7 was longer than that of AF7p, as confirmed by ex vivo imaging of the main organs. These results suggest the feasibility of using the subtraction biopanning strategy to screen the affinity peptide targeting MT-loop regions and HS7 is a superior probe for noninvasively imaging MT1-MMP expression in MT1-MMP-positive tumor models. It provides impetus for further studies to use HS7 in early diagnosis of tumors and in peptide-mediated drugs.

更新日期：2019-04-15

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