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Generation and Characterization of Novel Diagnostic Mouse Monoclonal Antibodies Against Human Estrogen Receptor Alpha and Progesterone Receptor Publisher Pubmed



Makoui MH1 ; Mobini M1 ; Khoshnoodi J1 ; Judaki MA1 ; Bahadori T1 ; Zare HA2 ; Golsazshirazi F1 ; Tabriz HM3 ; Madjd Z4 ; Jedditehrani M2 ; Zarnani AH1 ; Amiri MM1 ; Shokri F1
Authors
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Authors Affiliations
  1. 1. Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
  2. 2. Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran
  3. 3. Department of Pathology, Tehran University of Medical Sciences, Tehran, Iran
  4. 4. Oncopathology Research Center, Iran University of Medical Sciences (IUMS), Tehran, Iran

Source: Asian Pacific Journal of Cancer Prevention Published:2022


Abstract

Background: Estrogen and progesterone regulate the growth and development of several human cells and tissues. Their corresponding receptors (ER and PR) are important diagnostic and prognostic indicators for cancers of the breast and reproductive organs. Immunohistochemical analysis of ER and PR is the current standard method for evaluating the expression of these receptors in different cancers. Nonetheless, there is a significant lack of reproducibility of IHC results in laboratories worldwide, necessitating to develop more sensitive and specific antibodies for ER and PR IHC staining. Methods: ER and PR-specific monoclonal antibodies (MAbs) were generated by immunizing mice with synthetic peptides from ERα and PR. The isotypes and affinity constants of the selected MAbs were determined, and their specificities were assessed by peptide-specific ELISA, IHC, Western-blot analysis, and flow cytometry. In addition, the reactivity of generated MAbs was compared with that of the commercially-available anti-ER and anti-PR antibodies in IHC using normal and cancerous tissue sections. Moreover, 200 breast cancer tissue samples were stained using the newly generated MAbs along with commercial antibodies by IHC, and the sensitivity, specificity and accuracy of our MAbs were evaluated. Results: Among different MAbs generated in this study, two anti-ER and one anti-PR MAbs specifically detected the target antigens in normal and cancerous tissues in IHC. Further analyses confirmed the specificity of the MAbs in Western blotting and flow cytometry using a panel of ER and PR positive cell lines. The sensitivity, specificity and accuracy calculated for clone 1B9 (anti-ER) were 92.3%, 94.8% and 93%, and for clone 3D6 (anti-PR) were 93.0%, 94.3% and 93.5%, respectively. Conclusion: Our novel anti-ER and PR MAbs could be considered as suitable tools for diagnostic and research purposes. © This work is licensed under a Creative Commons Attribution-Non Commercial 4.0 International License