|تعداد مشاهده مقاله||12,897,021|
|تعداد دریافت فایل اصل مقاله||6,145,343|
Novel Monoclonal Antibodies Specific for Human Ki67 and P53 Tumor Markers in Breast Cancer Tissue Samples
|Iranian Journal of Immunology|
|مقاله 4، دوره 20، شماره 2، شهریور 2023، صفحه 177-189 اصل مقاله (3.8 M)|
|نوع مقاله: Original Article|
|شناسه دیجیتال (DOI): 10.22034/iji.2023.96926.2469|
|Masoud Hassanzadeh Makoui1؛ Maryam Mobini1؛ Jalal Khoshnoodi1؛ Tannaz Bahadori1؛ Forough Golsaz-Shirazi1؛ Hedieh Moradi Tabriz2؛ Zahra Madjd3؛ Mahmood Jeddi-Tehrani4؛ Amir-Hassan Zarnani1؛ Mohammad Mehdi Amiri* 1؛ Fazel Shokri* 1|
|1Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran|
|2Department of Pathology, Tehran University of Medical Sciences, Tehran, Iran|
|3Department of Pathology, Iran University of Medical Sciences, Tehran, Iran|
|4Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran|
|Background: Ki67 and P53 are important diagnostic and prognostic biomarkers expressed in several cancers. The current standard method for evaluating Ki67 and P53 in cancer tissues is immunohistochemistry (IHC), and having highly sensitive monoclonal antibodies against these biomarkers is necessary for an accurate diagnosis in the IHC test.|
Objective: To generate and characterize novel monoclonal antibodies (mAbs) against human Ki67 and P53 antigens for IHC purposes.
Methods: Ki67 and P53-specific mAbs were produced by the hybridoma method and screened by enzyme-linked immunosorbent assay (ELISA) and IHC techniques. Selected mAbs were characterized using Western blot and flow cytometry, and their affinities and isotypes were determined by ELISA. Moreover, using the IHC technique in 200 breast cancer tissue samples, we assessed the specificity, sensitivity, and accuracy of the produced mAbs.
Results: Two anti-Ki67 (2C2 and 2H1) and three anti-P53 mAbs (2A6, 2G4, and 1G10) showed strong reactivity to their target antigens in IHC. The selected mAbs were also able to recognize their targets by flow cytometry as well as Western blotting using human tumor cell lines expressing these antigens. The specificity, sensitivity, and accuracy calculated for clone 2H1 were 94.2%, 99.0%, and 96.6%, and for clone 2A6 were 97.3%, 98.1%, and 97.5%, respectively. Using these two monoclonal antibodies, we found a significant correlation between Ki67 and P53 overexpression and lymph node metastasis in patients with breast cancer.
Conclusion: The present study showed that the novel anti-Ki67 and anti-P53 mAbs could recognize their respective antigens with high specificity and sensitivity and therefore can be used in prognostic studies.
|Immunohistochemistry؛ Ki67؛ Monoclonal Antibody؛ P53|
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