Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and es...Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and established a rapid and accurate lateral flow immunoassay(LFIA)for use in community medical institutions.The gene sequence of human HER2-ECD was obtained from the National Center for Biotechnology Information(NCBI)to construct the expression plasmid.HER2-ECD protein expressed in HEK293F cells was used to immunize BALB/c mice.The monoclonal antibodies were produced in mouse ascites and isolated by hybridoma cell screening.Antibodies were analyzed for purity by SDS-PAGE(sodium dodecyl sulphate-polyacrylamide gel-electrophoresis)and affinity was assessed by enzyme-linked immunosorbent assay(ELISA)while subtypes were detected using the commercial kits.The HER2-ECD test strip was prepared based on the sandwich method and evaluated using a portable detection instrument.The affinity of the paired antibodies,4D8 and 8D9,both reached 1×108 L/mol.Both antibodies specifically recognized the HER2-ECD protein in serum.The limit of detection(LOD)of the gold nanoparticle(AuNP)-based LFIA was 1.7 ng/mL with a detection range of 1.7-400 ng/mL,and the performance of the HER2-ECD strip correlated well with that of a Siemens chemiluminescent immunoassay(CLIA)kit.In conclusion,the paired antibodies were successfully prepared with high affinity and specificity.The AuNP-based LFIA of HER2-ECD provides a fast and accurate method to detect the concentration of HER2-ECD in serum samples for clinical use in community medical institutions,and could contribute to determining the progress of the disease or the effectiveness of treatment.展开更多
Background: Head and neck squamous cell carcinoma (HNSCC), is a common malignancy in the Indian Population. In locally advanced disease, chemoradiation is the standard of care. Although induction chemotherapy has been...Background: Head and neck squamous cell carcinoma (HNSCC), is a common malignancy in the Indian Population. In locally advanced disease, chemoradiation is the standard of care. Although induction chemotherapy has been much studied, no clear benefit has been identified apart from laryngeal preservation. A few randomized trials have demonstrated improved response rate, disease free survival, and overall survival, with induction chemotherapy. Nimotuzumab is a humanized monoclonal antibody targeting epidermal growth factor receptors (EGFR). Unlike other Anti-EGFR monoclonal antibodies, it is demonstrated to be safer when combined with chemotherapy and/or radiotherapy. Aim: To evaluate the safety and efficacy of concurrently administered nimotuzumab with chemo-radiotherapy in patients with HNSCC in usual health care setting. Methods: This was an open-label, single arm study, with retrospective analysis of results. Patients above 18 years of age, and having histologically confirmed, advanced HNSCC were included in the study. The patients were treated with 3 cycles of induction chemotherapy consisting of modified TPF regimen along with nimotuzumab (200 mg IV) on Day 1, followed by radiotherapy for a dose of 66 Gy along with concurrent weekly cisplatin (30 mg/m2) and nimotuzumab (200 mg) throughout the course of radiation. Patients were evaluated using RECIST criteria, 4 weeks after the last cycle of chemotherapy. Results: Sixteen patients were included in this study, with mean age of 54 ± 11 years.?Most common sub-site of cancer was oral cavity in 69% (n = 11), followed by pharynx in 19% (n = 3).?Four patients had metastasis at the time of presentation. Six patients (37.5%) had progressive disease and four patients (25%) were lost to follow-up. The combination chemotherapy with nimotuzumab was well tolerated. Addition of nimotuzumab to TPF regimen was not associated with added toxicity. Conclusion: Addition of anti-EGFR monocloncal antibody (nimotuzumab) to induction chemotherapy and chemoradiation may be a promising alternative to concurrent chemoradiotherapy in HNSCC due to known over expression of EGFR receptors. The results of this study need further evaluation in a larger study setting.展开更多
基金the National Natural Science Foundation of China(No.22236002)National Key R&D Program(Nos.2023YFF1105003 and 2022YFA1207300).
文摘Human epidermal growth factor receptor 2(HER2)is an important biomarker for detection and treatment of breast cancer.In this study,we developed monoclonal antibodies against the extracellular domain(ECD)of HER2 and established a rapid and accurate lateral flow immunoassay(LFIA)for use in community medical institutions.The gene sequence of human HER2-ECD was obtained from the National Center for Biotechnology Information(NCBI)to construct the expression plasmid.HER2-ECD protein expressed in HEK293F cells was used to immunize BALB/c mice.The monoclonal antibodies were produced in mouse ascites and isolated by hybridoma cell screening.Antibodies were analyzed for purity by SDS-PAGE(sodium dodecyl sulphate-polyacrylamide gel-electrophoresis)and affinity was assessed by enzyme-linked immunosorbent assay(ELISA)while subtypes were detected using the commercial kits.The HER2-ECD test strip was prepared based on the sandwich method and evaluated using a portable detection instrument.The affinity of the paired antibodies,4D8 and 8D9,both reached 1×108 L/mol.Both antibodies specifically recognized the HER2-ECD protein in serum.The limit of detection(LOD)of the gold nanoparticle(AuNP)-based LFIA was 1.7 ng/mL with a detection range of 1.7-400 ng/mL,and the performance of the HER2-ECD strip correlated well with that of a Siemens chemiluminescent immunoassay(CLIA)kit.In conclusion,the paired antibodies were successfully prepared with high affinity and specificity.The AuNP-based LFIA of HER2-ECD provides a fast and accurate method to detect the concentration of HER2-ECD in serum samples for clinical use in community medical institutions,and could contribute to determining the progress of the disease or the effectiveness of treatment.
文摘Background: Head and neck squamous cell carcinoma (HNSCC), is a common malignancy in the Indian Population. In locally advanced disease, chemoradiation is the standard of care. Although induction chemotherapy has been much studied, no clear benefit has been identified apart from laryngeal preservation. A few randomized trials have demonstrated improved response rate, disease free survival, and overall survival, with induction chemotherapy. Nimotuzumab is a humanized monoclonal antibody targeting epidermal growth factor receptors (EGFR). Unlike other Anti-EGFR monoclonal antibodies, it is demonstrated to be safer when combined with chemotherapy and/or radiotherapy. Aim: To evaluate the safety and efficacy of concurrently administered nimotuzumab with chemo-radiotherapy in patients with HNSCC in usual health care setting. Methods: This was an open-label, single arm study, with retrospective analysis of results. Patients above 18 years of age, and having histologically confirmed, advanced HNSCC were included in the study. The patients were treated with 3 cycles of induction chemotherapy consisting of modified TPF regimen along with nimotuzumab (200 mg IV) on Day 1, followed by radiotherapy for a dose of 66 Gy along with concurrent weekly cisplatin (30 mg/m2) and nimotuzumab (200 mg) throughout the course of radiation. Patients were evaluated using RECIST criteria, 4 weeks after the last cycle of chemotherapy. Results: Sixteen patients were included in this study, with mean age of 54 ± 11 years.?Most common sub-site of cancer was oral cavity in 69% (n = 11), followed by pharynx in 19% (n = 3).?Four patients had metastasis at the time of presentation. Six patients (37.5%) had progressive disease and four patients (25%) were lost to follow-up. The combination chemotherapy with nimotuzumab was well tolerated. Addition of nimotuzumab to TPF regimen was not associated with added toxicity. Conclusion: Addition of anti-EGFR monocloncal antibody (nimotuzumab) to induction chemotherapy and chemoradiation may be a promising alternative to concurrent chemoradiotherapy in HNSCC due to known over expression of EGFR receptors. The results of this study need further evaluation in a larger study setting.
