Use of photoimmunoconjugates to characterize ABCB1 in cancer cells.
Use of photoimmunoconjugates to characterize ABCB1 in cancer cells.
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使用光免疫缀合物表征癌细胞中的ABCB 1。
DOI：
10.1515/nanoph-2021-0252
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发表时间：
2021-09
期刊：
影响因子：
7.5
通讯作者：
Huang HC
中科院分区：
文献类型：
--
作者：
Liang BJ;Lusvarghi S;Ambudkar SV;Huang HC
准确检测ATP结合盒药物转运体ABCB1的表达对于准确识别耐药肿瘤是必不可少的。现有的检测方法不能提供关于转运体功能状态的必要的分子细节。光免疫结合物是一类独特的抗体-染料结合物，用于分子诊断和治疗。然而，将疏水性光敏剂与亲水性抗体偶联是相当具有挑战性的。在这里，我们设计了一种光免疫结合物，它结合了临床批准的苯并卟啉衍生物(BPD)光敏剂和构象敏感的UIC2单抗来靶向功能活性的人ABCB1(即，内向开放构象的ABCB1)。我们发现UIC2的聚乙二醇化提高了BPD的共轭效率，减少了17%的非共价共轭BPD分子的数量。尺寸排除层析有效地分离了UIC2-BPD样品中发现的不同分子量物种。在脂类纳米盘和高表达ABCB1的三阴性乳腺癌(TNBC)细胞中证实了UIC2-BPD与ABCB1的结合。UIC2-BPD被发现保持了UIC2的构象敏感性，因为ABCB1调节剂的加入提高了体外抗体的反应性。因此，BPD的固有荧光能力可用于通过UIC2-BPD标记ABCB1过表达的TNBC细胞。我们的发现为疏水性光敏剂与针对癌细胞表面表达的靶蛋白的构象敏感抗体的结合提供了洞察力。
Accurate detection of ATP-binding cassette drug transporter ABCB1 expression is imperative for precise identification of drug-resistant tumors. Existing detection methods fail to provide the necessary molecular details regarding the functional state of the transporter. Photo-immunoconjugates are a unique class of antibody–dye conjugates for molecular diagnosis and therapeutic treatment. However, conjugating hydrophobic photosensitizers to hydrophilic antibodies is quite challenging. Here, we devise a photoimmunoconjugate that combines a clinically approved benzoporphyrin derivative (BPD) photosensitizer and the conformational-sensitive UIC2 monoclonal antibody to target functionally active human ABCB1 (i.e., ABCB1 in the inward-open conformation). We show that PEGylation of UIC2 enhances the BPD conjugation efficiency and reduces the amount of non-covalently conjugated BPD molecules by 17%. Size exclusion chromatography effectively separates the different molecular weight species found in the UIC2–BPD sample. The binding of UIC2–BPD to ABCB1 was demonstrated in lipidic nanodiscs and ABCB1-overexpressing triple negative breast cancer (TNBC) cells. UIC2–BPD was found to retain the conformation sensitivity of UIC2, as the addition of ABCB1 modulators increases the antibody reactivity in vitro. Thus, the inherent fluorescence capability of BPD can be used to label ABCB1-overexpressing TNBC cells using UIC2–BPD. Our findings provide insight into conjugation of hydrophobic photosensitizers to conformation-sensitive antibodies to target proteins expressed on the surface of cancer cells.
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影响因子：
4.9
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