The generation of the recombinant human MSLN protein typically includes the following steps: synthesizing the recombinant plasmid containing the gene encoding the human MSLN protein (296-580aa) along with the N-terminal hFc-tag gene, transforming the recombinant plasmid into mammalian cells, selecting the positive mammalian cells, and culturing the positive mammalian cells for protein expression. After expression, affinity purification is employed to isolate and purify the recombinant MSLN protein from the cell lysate. Denaturing SDS-PAGE is utilized to resolve the resulting recombinant human MSLN protein, revealing a purity level exceeding 90%. The endotoxin of this protein is less than 1.0 EU/ug as determined by the LAL method. Its activity has been validated through the functional ELISA.
Mesothelin (MSLN) is a protein found on the surface of certain cancer cells, including those from mesothelioma, ovarian cancer, and pancreatic cancer [1]. It's often used as a marker to identify these types of tumors [2]. The gene that makes MSLN produces a larger protein that gets split into two smaller ones: Megakaryocytic Potentiating Factor (MPF) and mature MSLN (mMSLN) [3]. In ovarian cancer, MSLN interacts with a protein called CA-125, possibly aiding in the spread of cancer cells [4]. While MSLN levels are high in ovarian cancer, their exact role in tumor growth isn't fully understood [5].
Researchers are studying MSLN as a potential marker and target for treatment in mesothelioma [6]. Some believe MSLN helps cancer cells stick to certain tissues and spread in ovarian cancer [7]. However, because MSLN levels vary in different types of tumors, it's unclear if it directly contributes to cancer development [8]. Still, studies suggest that MSLN might spur cell growth in colorectal cancer and encourage the invasion of mesothelioma cells, indicating it could play a role in cancer progression [6]. Additionally, MSLN seems to influence cell growth in pancreatic cancer by affecting certain proteins involved in cell division [9].
References:
[1] S. Inoue, T. Tsunoda, M. Riku, H. Ito, A. Inoko, H. Murakamiet al., Diffuse mesothelin expression leads to worse prognosis through enhanced cellular proliferation in colorectal cancer, Oncology Letters, 2020. https://doi.org/10.3892/ol.2020.11290
[2] X. Hou, F. Wang, X. Meng, D. Li, J. Ding, Y. Chenet al., Construction of a 124i-labeled specific antibody for the noninvasive detection of mesothelin-overexpressing tumors, Molecular Pharmaceutics, vol. 19, no. 10, p. 3623-3631, 2022. https://doi.org/10.1021/acs.molpharmaceut.2c00342
[3] J. Zhang, T. Bera, W. Liu, X. Du, C. Alewine, R. Hassanet al., Megakaryocytic potentiating factor and mature mesothelin stimulate the growth of a lung cancer cell line in the peritoneal cavity of mice, Plos One, vol. 9, no. 8, p. e104388, 2014. https://doi.org/10.1371/journal.pone.0104388
[4] T. Hilliard, The impact of mesothelin in the ovarian cancer tumor microenvironment, Cancers, vol. 10, no. 9, p. 277, 2018. https://doi.org/10.3390/cancers10090277
[5] T. Hilliard, B. Kowalski, K. Iwamoto, E. Agadi, Y. Liu, J. Yanget al., Host mesothelin expression increases ovarian cancer metastasis in the peritoneal microenvironment, International Journal of Molecular Sciences, vol. 22, no. 22, p. 12443, 2021. https://doi.org/10.3390/ijms222212443
[6] E. Servais, C. Colovos, L. Rodríguez, A. Bograd, J. Nitadori, C. Simaet al., Mesothelin overexpression promotes mesothelioma cell invasion and mmp-9 secretion in an orthotopic mouse model and in epithelioid pleural mesothelioma patients, Clinical Cancer Research, vol. 18, no. 9, p. 2478-2489, 2012. https://doi.org/10.1158/1078-0432.ccr-11-2614
[7] B. Wang, J. Kuroiwa, L. He, A. Charalambous, T. Keler, & R. Steinman, The human cancer antigen mesothelin is more efficiently presented to the mouse immune system when targeted to the dec‐205/cd205 receptor on dendritic cells, Annals of the New York Academy of Sciences, vol. 1174, no. 1, p. 6-17, 2009. https://doi.org/10.1111/j.1749-6632.2009.04933.x
[8] X. He, E. Despeaux, T. Stueckle, A. Chi, V. Castranova, C. Dinuet al., Role of mesothelin in carbon nanotube-induced carcinogenic transformation of human bronchial epithelial cells, Ajp Lung Cellular and Molecular Physiology, vol. 311, no. 3, p. L538-L549, 2016. https://doi.org/10.1152/ajplung.00139.2016
[9] U. Bharadwaj, M. Li, C. Chen, & Q. Yao, Mesothelin-induced pancreatic cancer cell proliferation involves alteration of cyclin e via activation of signal transducer and activator of transcription protein 3, Molecular Cancer Research, vol. 6, no. 11, p. 1755-1765, 2008. https://doi.org/10.1158/1541-7786.mcr-08-0095
