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The synthesis of the recombinant human CD70 protein involves gene cloning, plasmid assembly, protein expression, purification, and analysis. The gene sequence encoding amino acids 52-193 of the human CD70 is co-inserted into a plasmid with an N-terminal 10xHis-tag gene. The plasmid is transfected into mammalian cells, and a selective antibiotic is introduced to screen the transfected cells. The selected cells are cultured for protein expression, and the CD70 protein is released by lysing the cells and then purified via Ni-NTA affinity chromatography. SDS-PAGE confirms a purity level of the recombinant CD70 greater than 92.5%, and the LAL method verifies endotoxin content below 1.0 EU/μg. The CD70 protein's activity is validated through ELISA, in which it binds to the CD70 antibody with an EC50 of 2.414-3.196 ng/mL.
The human CD70 antigen is a member of the TNF superfamily, primarily expressed on activated lymphocytes and certain tumor cells. CD70 interacts with its receptor CD27, which is crucial for T-cell activation, proliferation, and survival, thereby influencing immune responses against tumors [3][10].
CD70 expression is notably upregulated in various malignancies, including renal cell carcinoma (RCC), melanoma, and glioblastoma, among others. Elevated expression of CD70 is often associated with aggressive tumor behavior and poor prognosis [6][8][9]. Studies have shown that CD70 is expressed in approximately 30% to 68% of clear cell RCC cases and is linked to hematogenous metastases in pancreatic cancer [1][8]. The presence of CD70 in these tumors suggests its involvement in promoting tumor growth and immune evasion, as it can induce apoptosis in T-cells and B-cells, thereby facilitating tumor progression [6].
CD70's restricted expression pattern—being largely absent in normal tissues—makes it an attractive target for immunotherapy. Several therapeutic strategies are being explored, including monoclonal antibodies and antibody-drug conjugates (ADCs) targeting CD70. These approaches aim to harness the immune system to selectively destroy CD70-expressing tumor cells while minimizing damage to normal cells [2][4][5]. For example, the humanized antibody SGN-70 has shown promising antitumor activity in preclinical models, highlighting the potential of CD70-targeted therapies in treating malignancies [7].
References:
[1] Flieswasser, T., Camara-Clayette, V., Danu, A., Bosq, J., Ribrag, V., Zabrocki, P., … & Jacobs, J. (2019). Screening a broad range of solid and haematological tumour types for cd70 expression using a uniform ihc methodology as potential patient stratification method. Cancers, 11(10), 1611. https://doi.org/10.3390/cancers11101611
[2] Hagemann, U., Mihaylova, D., Uran, S., Borrebaek, J., Grant, D., Bjerke, R., … & Cuthbertson, A. (2017). Targeted alpha therapy using a novel cd70 targeted thorium-227 conjugate inin vitroandin vivomodels of renal cell carcinoma. Oncotarget, 8(34), 56311-56326. https://doi.org/10.18632/oncotarget.16910
[3] Jacobs, J., Zwaenepoel, K., Rolfo, C., Bossche, J., Deben, C., Silence, K., … & Pauwels, P. (2015). Unlocking the potential of cd70 as a novel immunotherapeutic target for non-small cell lung cancer. Oncotarget, 6(15), 13462-13475. https://doi.org/10.18632/oncotarget.3880
[4] Jeffrey, S., Burke, P., Lyon, R., Meyer, D., Sussman, D., Anderson, M., … & Senter, P. (2013). A potent anti-cd70 antibody–drug conjugate combining a dimeric pyrrolobenzodiazepine drug with site-specific conjugation technology. Bioconjugate Chemistry, 24(7), 1256-1263. https://doi.org/10.1021/bc400217g
[5] Jilaveanu, L., Sznol, J., Aziz, S., Duchen, D., Kluger, H., & Camp, R. (2012). Cd70 expression patterns in renal cell carcinoma. Human Pathology, 43(9), 1394-1399. https://doi.org/10.1016/j.humpath.2011.10.014
[6] Liu, N., Sheng, X., Liu, Y., & Yu, J. (2013). Increased cd70 expression is associated with clinical resistance to cisplatin-based chemotherapy and poor survival in advanced ovarian carcinomas. Oncotargets and Therapy, 615. https://doi.org/10.2147/ott.s44445
[7] McEarchern, J., Smith, L., McDonagh, C., Klussman, K., Gordon, K., Morris-Tilden, C., … & Law, C. (2008). Preclinical characterization of sgn-70, a humanized antibody directed against cd70. Clinical Cancer Research, 14(23), 7763-7772. https://doi.org/10.1158/1078-0432.ccr-08-0493
[8] Nakamura, K., Sho, M., Akahori, T., Nishiwada, S., Kunishige, T., Nakagawa, K., … & Ikeda, N. (2021). Clinical relevance of cd70 expression in resected pancreatic cancer: prognostic value and therapeutic potential. Pancreatology, 21(3), 573-580. https://doi.org/10.1016/j.pan.2021.01.013
[9] Pich-Bavastro, C., Sarrabayrouse, G., Teiti, I., Mariamé, B., Rochaix, P., Lamant, L., … & Tilkin–Mariamé, A. (2015). Melanoma-expressed cd70 is involved in invasion and metastasis. British Journal of Cancer, 114(1), 63-70. https://doi.org/10.1038/bjc.2015.412
[10] Shaffer, D., Savoldo, B., Yi, Z., Chow, K., Kakarla, S., Spencer, D., … & Gottschalk, S. (2011). T cells redirected against cd70 for the immunotherapy of cd70-positive malignancies. Blood, 117(16), 4304-4314. https://doi.org/10.1182/blood-2010-04-278218
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