PDF(Pubmed)
Abstract：
OBJECTIVE: Breast cancer (BC) is the most common malignant disease worldwide. Localized stages of BC can be successfully treated by surgery. However, local recurrence occurs in about 4-10% of patients, requiring systemic treatments that impair the patients\' quality of life and shortens life expectancy. Therefore, new therapeutic options are needed, which can be used intraoperatively and contribute to the complete removal of residual tumor cells in the surgical area. In the present study, we describe a cysteine-modified variant of the anti-HER2 antibody trastuzumab, that was coupled to the silicon phthalocyanine photosensitizer dye WB692-CB1 for the photoimmunotherapy (PIT) of BC.
METHODS: The cysteine modified trastuzumab variant was cloned and expressed in Expi293F cells. After purification via immobilized affinity chromatography, the antibody was coupled to the dye. Cell binding of the antibody and the antibody dye conjugate was measured by flow cytometry. After incubation of BC cells with the conjugate and activation of the dye by irradiation with red light, cell viability was determined.
RESULTS: The antibody and the conjugate showed specific binding to HER2-expressing BC cells. Treatment of the HER2high BC cell line SK-BR-3 with the conjugate followed by irradiation with a red light dose of 32 J/cm2 led to complete cell killing within 24 h.
CONCLUSIONS: Our novel antibody dye conjugate represents a promising candidate for intraoperative treatment of localized BC, aiming to eliminate residual tumor cells in the surgical area and potentially reduce local recurrence, thereby improving recovery prospects for BC patients.
METHODS: The cysteine modified trastuzumab variant was cloned and expressed in Expi293F cells. After purification via immobilized affinity chromatography, the antibody was coupled to the dye. Cell binding of the antibody and the antibody dye conjugate was measured by flow cytometry. After incubation of BC cells with the conjugate and activation of the dye by irradiation with red light, cell viability was determined.
RESULTS: The antibody and the conjugate showed specific binding to HER2-expressing BC cells. Treatment of the HER2high BC cell line SK-BR-3 with the conjugate followed by irradiation with a red light dose of 32 J/cm2 led to complete cell killing within 24 h.
CONCLUSIONS: Our novel antibody dye conjugate represents a promising candidate for intraoperative treatment of localized BC, aiming to eliminate residual tumor cells in the surgical area and potentially reduce local recurrence, thereby improving recovery prospects for BC patients.
摘要：
目的：乳腺癌（BC）是全球最常见的恶性疾病。局部阶段的BC可以通过手术成功治疗。然而,局部复发发生在约4-10%的患者，需要系统性治疗，损害患者的生活质量，缩短预期寿命。因此,需要新的治疗选择，可以在术中使用，并有助于完全去除手术区域的残留肿瘤细胞。在本研究中,我们描述了抗HER2抗体曲妥珠单抗的半胱氨酸修饰变体，与硅酞菁光敏剂染料WB692-CB1偶联，用于BC的光免疫疗法（PIT）。
方法:克隆半胱氨酸修饰的曲妥珠单抗变体并在Expi293F细胞中表达。通过固定化亲和层析纯化后，抗体与染料偶联。通过流式细胞术测量抗体和抗体染料缀合物的细胞结合。将BC细胞与缀合物孵育并通过红光照射激活染料后，确定细胞活力。
结果:抗体和缀合物显示与表达HER2的BC细胞的特异性结合。用缀合物处理HER2highBC细胞系SK-BR-3，然后用32J/cm2的红光剂量照射导致24小时内完全杀死细胞。
结论：我们的新型抗体染料缀合物代表了局部BC术中治疗的有希望的候选者，旨在消除手术区域残留的肿瘤细胞，并有可能减少局部复发，从而改善BC患者的康复前景。
方法:克隆半胱氨酸修饰的曲妥珠单抗变体并在Expi293F细胞中表达。通过固定化亲和层析纯化后，抗体与染料偶联。通过流式细胞术测量抗体和抗体染料缀合物的细胞结合。将BC细胞与缀合物孵育并通过红光照射激活染料后，确定细胞活力。
结果:抗体和缀合物显示与表达HER2的BC细胞的特异性结合。用缀合物处理HER2highBC细胞系SK-BR-3，然后用32J/cm2的红光剂量照射导致24小时内完全杀死细胞。
结论：我们的新型抗体染料缀合物代表了局部BC术中治疗的有希望的候选者，旨在消除手术区域残留的肿瘤细胞，并有可能减少局部复发，从而改善BC患者的康复前景。
