{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T22:44:23Z","timestamp":1790030663275,"version":"4.0.1"},"reference-count":55,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2022,3,16]],"date-time":"2022-03-16T00:00:00Z","timestamp":1647388800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Center","doi-asserted-by":"publisher","award":["2018\/31\/N\/NZ6\/03004"],"award-info":[{"award-number":["2018\/31\/N\/NZ6\/03004"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Approximately one-third of the human population is infected with the intracellular cosmopolitan protozoan Toxoplasma gondii (Tg), and a specific treatment for this parasite is still needed. Additionally, the increasing resistance of Tg to drugs has become a challenge for numerous research centers. The high selectivity of a compound toward the protozoan, along with low cytotoxicity toward the host cells, form the basis for further research, which aims at determining the molecular targets of the active compounds. Thiosemicarbazide derivatives are biologically active organic compounds. Previous studies on the initial preselection of 58 new 4-arylthiosemicarbazide derivatives in terms of their anti-Tg activity and selectivity made it possible to select two promising derivatives for further research. One of the important amino acids involved in the proliferation of Tg and the formation of parasitophorous vacuoles is tyrosine, which is converted by two unique aromatic amino acid hydroxylases to levodopa. Enzymatic studies with two derivatives (R: para-nitro and meta-iodo) and recombinant aromatic amino acid hydroxylase (AAHs) obtained in the E. coli expression system were performed, and the results indicated that toxoplasmic AAHs are a molecular target for 4-arylthiosemicarbazide derivatives. Moreover, the drug affinity responsive target stability assay also confirmed that the selected compounds bind to AAHs. Additionally, the anti-inflammatory activity of these derivatives was tested using THP1-Blue\u2122 NF-\u03baB reporter cells due to the similarity of the thiosemicarbazide scaffold to thiosemicarbazone, both of which are known NF-\u03baB pathway inhibitors.<\/jats:p>","DOI":"10.3390\/ijms23063213","type":"journal-article","created":{"date-parts":[[2022,3,16]],"date-time":"2022-03-16T22:09:58Z","timestamp":1647468598000},"page":"3213","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["4-Arylthiosemicarbazide Derivatives as Toxoplasmic Aromatic Amino Acid Hydroxylase Inhibitors and Anti-inflammatory Agents"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4980-1074","authenticated-orcid":false,"given":"Adrian","family":"Bekier","sequence":"first","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7335-9853","authenticated-orcid":false,"given":"Anna","family":"Brzostek","sequence":"additional","affiliation":[{"name":"Laboratory of Genetics and Physiology of Mycobacterium, Institute of Medical Biology, Polish Academy of Sciences, 93-232 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Agata","family":"Paneth","sequence":"additional","affiliation":[{"name":"Department of Organic Chemistry, Faculty of Pharmacy, Medical University of Lublin, 20-093 Lublin, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4942-7385","authenticated-orcid":false,"given":"Bo\u017cena","family":"Dziadek","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1427-884X","authenticated-orcid":false,"given":"Jaros\u0142aw","family":"Dziadek","sequence":"additional","affiliation":[{"name":"Laboratory of Genetics and Physiology of Mycobacterium, Institute of Medical Biology, Polish Academy of Sciences, 93-232 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0559-1706","authenticated-orcid":false,"given":"Justyna","family":"Gatkowska","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3265-0409","authenticated-orcid":false,"given":"Katarzyna","family":"Dzitko","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology, Faculty of Biology and Environmental Protection, University of Lodz, 90-237 Lodz, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,3,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1965","DOI":"10.1016\/S0140-6736(04)16412-X","article-title":"Toxoplasmosis","volume":"363","author":"Montoya","year":"2004","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"e00054","DOI":"10.1016\/j.fawpar.2019.e00054","article-title":"Toxoplasmosis: Overview from a One Health perspective","volume":"15","author":"Dubey","year":"2019","journal-title":"Food Waterborne Parasitol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S1473-3099(14)70772-8","article-title":"A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for cryptosporidium","volume":"15","author":"Checkley","year":"2015","journal-title":"Lancet Infect. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/B978-0-12-397900-1.00001-3","article-title":"The global public health significance of Plasmodium vivax","volume":"80","author":"Battle","year":"2012","journal-title":"Adv. Parasitol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Havelaar, A.H., Kirk, M.D., Torgerson, P.R., Gibb, H.J., Hald, T., Lake, R.J., Praet, N., Bellinger, D.C., de Silva, N.R., and Gargouri, N. (2015). World Health Organization Global Estimates and Regional Comparisons of the Burden of Foodborne Disease in 2010. PLoS Med., 12.","DOI":"10.1371\/journal.pmed.1001923"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"501","DOI":"10.2471\/BLT.12.111732","article-title":"The global burden of congenital toxoplasmosis: A systematic review","volume":"91","author":"Torgerson","year":"2013","journal-title":"Bull. World Health Organ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e00037","DOI":"10.1016\/j.fawpar.2019.e00037","article-title":"infection and toxoplasmosis in farm animals: Risk factors and economic impact","volume":"15","author":"Stelzer","year":"2019","journal-title":"Food Waterborne Parasitol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1217","DOI":"10.1016\/S0020-7519(00)00124-7","article-title":"Toxoplasma gondii: From animals to humans","volume":"30","author":"Tenter","year":"2000","journal-title":"Int. J. Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Krishnan, A., and Soldati-Favre, D. (2021). Amino Acid Metabolism in Apicomplexan Parasites. Metabolites, 11.","DOI":"10.3390\/metabo11020061"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1719","DOI":"10.12688\/f1000research.16021.1","article-title":"Metabolic interactions between Toxoplasma gondii and its host","volume":"7","author":"Blume","year":"2018","journal-title":"F1000Research"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Parker, K.E.R., Fairweather, S.J., Rajendran, E., Blume, M., McConville, M.J., Br\u00f6er, S., Kirk, K., and van Dooren, G.G. (2019). The tyrosine transporter of Toxoplasma gondii is a member of the newly defined apicomplexan amino acid transporter (ApiAT) family. PLoS Pathog., 15.","DOI":"10.1371\/journal.ppat.1007577"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.exppara.2017.02.018","article-title":"Toxoplasma growth in vitro is dependent on exogenous tyrosine and is independent of AAH2 even in tyrosine-limiting conditions","volume":"176","author":"Marino","year":"2017","journal-title":"Exp. Parasitol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Wang, Z.T., Verma, S.K., Dubey, J.P., and Sibley, L.D. (2017). The aromatic amino acid hydroxylase genes AAH1 and AAH2 in Toxoplasma gondii contribute to transmission in the cat. PLoS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006272"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Gaskell, E.A., Smith, J.E., Pinney, J.W., Westhead, D.R., and McConkey, G.A. (2009). A unique dual activity amino acid hydroxylase in Toxoplasma gondii. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004801"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1128\/EC.2.3.456-464.2003","article-title":"Roles of tyrosine-rich precursor glycoproteins and dityrosine- and 3,4-dihydroxyphenylalanine-mediated protein cross-linking in development of the oocyst wall in the coccidian parasite Eimeria maxima","volume":"2","author":"Belli","year":"2003","journal-title":"Eukaryot. Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1080\/14756366.2021.1931164","article-title":"4-Arylthiosemicarbazide derivatives as a new class of tyrosinase inhibitors and anti-Toxoplasma gondii agents","volume":"36","author":"Bekier","year":"2021","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Zhang, Z., Li, Y., Li, H., Song, X., Ma, Z., Lu, H., Liu, S., Zhao, Y., Tan, M., and Wang, S. (2020). Identification of Toxoplasma gondii tyrosine hydroxylase (TH) activity and molecular immunoprotection against toxoplasmosis. Vaccines, 8.","DOI":"10.3390\/vaccines8020158"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/j.bbr.2018.03.023","article-title":"AAH2 gene is not required for dopamine-dependent neurochemical and behavioral abnormalities produced by Toxoplasma infection in mouse","volume":"347","author":"McFarland","year":"2018","journal-title":"Behav. Brain Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"13822","DOI":"10.1038\/s41598-017-13229-y","article-title":"Toxoplasma-induced changes in host risk behaviour are independent of parasite-derived AaaH2 tyrosine hydroxylase","volume":"7","author":"Afonso","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Wei, H.X., Wei, S.S., Lindsay, D.S., and Peng, H.J. (2015). A Systematic Review and Meta-Analysis of the Efficacy of Anti-Toxoplasma gondii Medicines in Humans. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0138204"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1007\/s40268-017-0206-8","article-title":"Adverse Event Profile of Pyrimethamine-Based Therapy in Toxoplasmosis: A Systematic Review","volume":"17","author":"Goodwin","year":"2017","journal-title":"Drugs RD"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Joynson, D.H.M., and Wreghitt, T.G. (2001). Toxoplasmosis: A Comprehensive Clinical Guide, Cambridge University Press.","DOI":"10.1017\/CBO9780511527005"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"9974","DOI":"10.1021\/acs.jmedchem.7b01663","article-title":"Novel Molecules to Treat Chronic Central Nervous System Toxoplasmosis","volume":"60","author":"Kortagere","year":"2017","journal-title":"J. Med. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.1021\/jm950760y","article-title":"Lipophilic antifolates as agents against opportunistic infections. 1. Agents superior to trimetrexate and piritrexim against Toxoplasma gondii and Pneumocystis carinii in in vitro evaluations","volume":"39","author":"Piper","year":"1996","journal-title":"J. Med. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2587","DOI":"10.3389\/fmicb.2018.02587","article-title":"Drug Resistance in Toxoplasma gondii","volume":"9","author":"Montazeri","year":"2018","journal-title":"Front. Microbiol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Mayers, D.L., Sobel, J.D., Ouellette, M., Kaye, K.S., and Marchaim, D. (2017). Antimicrobial Drug Resistance: Mechanisms of Drug Resistance, Springer International Publishing. [2nd ed.].","DOI":"10.1007\/978-3-319-46718-4"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"925","DOI":"10.1016\/j.ejmech.2012.06.003","article-title":"Synthesis and antitumor activity of novel quinazoline derivatives containing thiosemicarbazide moiety","volume":"54","author":"He","year":"2012","journal-title":"Eur. J. Med. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"488","DOI":"10.1016\/j.foodchem.2013.01.027","article-title":"4-Methyl-7-hydroxycoumarin antifungal and antioxidant activity enhancement by substitution with thiosemicarbazide and thiazolidinone moieties","volume":"139","author":"Molnar","year":"2013","journal-title":"Food Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"11520","DOI":"10.3390\/molecules190811520","article-title":"New thiosemicarbazides and 1,2,4-triazolethiones derived from 2-(ethylsulfanyl) benzohydrazide as potent antioxidants","volume":"19","author":"Nazarbahjat","year":"2014","journal-title":"Molecules"},{"key":"ref_30","first-page":"1128","article-title":"Synthesis and Biological Evaluation of Thiosemicarbazide Derivatives Endowed with High Activity toward Mycobacterium bovis","volume":"16","author":"Sardari","year":"2017","journal-title":"Iran J. Pharm. Res."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Chudzik-Rz\u0105d, B., Malm, A., Trotsko, N., Wujec, M., Plech, T., and Paneth, A. (2020). Synergistic Effects of Thiosemicarbazides with Clinical Drugs against S. aureus. Molecules, 25.","DOI":"10.3390\/molecules25102302"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Bekier, A., Kawka, M., Lach, J., Dziadek, J., Paneth, A., Gatkowska, J., Dzitko, K., and Dziadek, B. (2021). Imidazole-Thiosemicarbazide Derivatives as Potent Anti-Mycobacterium tuberculosis Compounds with Antibiofilm Activity. Cells, 10.","DOI":"10.3390\/cells10123476"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"107778","DOI":"10.1016\/j.intimp.2021.107778","article-title":"Therapeutic perspective of thiosemicarbazones derivatives in inflammatory pathologies: A summary of in vitro\/in vivo studies","volume":"96","author":"Kanso","year":"2021","journal-title":"Int. Immunopharmacol."},{"key":"ref_34","first-page":"15","article-title":"Thiosemicarbazone and thiazolylhydrazones of 1-indanone: As a new class of nonacidic anti-inflammatory and antiplatelet aggregation agents","volume":"32","author":"Nazim","year":"2019","journal-title":"Pak. J. Pharm. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1007\/s43440-021-00221-7","article-title":"Anti-inflammatory activity of novel thiosemicarbazone compounds indole-based as COX inhibitors","volume":"73","author":"Jacob","year":"2021","journal-title":"Pharmacol. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Shawish, H.B., Wong, W.Y., Wong, Y.L., Loh, S.W., Looi, C.Y., Hassandarvish, P., Phan, A.Y., Wong, W.F., Wang, H., and Paterson, I.C. (2014). Nickel(II) complex of polyhydroxybenzaldehyde N4-thiosemicarbazone exhibits anti-inflammatory activity by inhibiting NF-\u03baB transactivation. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100933"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4359","DOI":"10.1242\/jcs.00683","article-title":"Activation of NF-kappaB by Toxoplasma gondii correlates with increased expression of antiapoptotic genes and localization of phosphorylated IkappaB to the parasitophorous vacuole membrane","volume":"116","author":"Molestina","year":"2003","journal-title":"J. Cell Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Paneth, A., W\u0119gli\u0144ska, L., Bekier, A., Stefaniszyn, E., Wujec, M., Trotsko, N., and Dzitko, K. (2019). Systematic Identification of Thiosemicarbazides for Inhibition of Toxoplasma gondii Growth In Vitro. Molecules, 24.","DOI":"10.3390\/molecules24030614"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Paneth, A., W\u0119gli\u0144ska, L., Bekier, A., Stefaniszyn, E., Wujec, M., Trotsko, N., Hawry\u0142, A., Hawry\u0142, M., and Dzitko, K. (2019). Discovery of Potent and Selective Halogen-Substituted Imidazole-Thiosemicarbazides for Inhibition of Toxoplasma gondii Growth In Vitro via Structure-Based Design. Molecules, 24.","DOI":"10.3390\/molecules24081618"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1080\/14756366.2018.1545767","article-title":"A comprehensive review on tyrosinase inhibitors","volume":"34","author":"Zolghadri","year":"2019","journal-title":"J. Enzyme Inhib. Med. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1016\/j.compbiolchem.2018.03.007","article-title":"In silico modelling of azole derivatives with tyrosinase inhibition ability: Application of the models for activity prediction of new compounds","volume":"74","author":"De","year":"2018","journal-title":"Comput. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1096","DOI":"10.1016\/j.bmc.2007.10.102","article-title":"1-(1-Arylethylidene)thiosemicarbazide derivatives: A new class of tyrosinase inhibitors","volume":"16","author":"Liu","year":"2008","journal-title":"Bioorg. Med. Chem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1016\/j.ejmech.2008.04.002","article-title":"A class of potent tyrosinase inhibitors: Alkylidenethiosemicarbazide compounds","volume":"44","author":"Liu","year":"2009","journal-title":"Eur. J. Med. Chem."},{"key":"ref_44","first-page":"e59656","article-title":"A Semi-Quantitative Drug Affinity Responsive Target Stability (DARTS) assay for studying Rapamycin\/mTOR interaction","volume":"150","author":"Zhang","year":"2019","journal-title":"J. Vis. Exp."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1084\/jem.20100717","article-title":"Strain-specific activation of the NF-kappaB pathway by GRA15, a novel Toxoplasma gondii dense granule protein","volume":"208","author":"Rosowski","year":"2011","journal-title":"J. Exp. Med."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e00808-19","DOI":"10.1128\/mBio.00808-19","article-title":"GRA15 Activates the NF-\u03baB Pathway through Interactions with TNF Receptor-Associated Factors","volume":"10","author":"Yang","year":"2019","journal-title":"mBio"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1084\/jem.184.2.597","article-title":"Association of CD4+ T cell-dependent, interferon-gamma-mediated necrosis of the small intestine with genetic susceptibility of mice to peroral infection with Toxoplasma gondii","volume":"184","author":"Liesenfeld","year":"1996","journal-title":"J. Exp. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"8785","DOI":"10.4049\/jimmunol.177.12.8785","article-title":"Gram-negative bacteria aggravate murine small intestinal Th1-type immunopathology following oral infection with Toxoplasma gondii","volume":"177","author":"Heimesaat","year":"2006","journal-title":"J. Immunol."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Hatter, J.A., Kouche, Y.M., Melchor, S.J., Ng, K., Bouley, D.M., Boothroyd, J.C., and Ewald, S.E. (2018). Toxoplasma gondii infection triggers chronic cachexia and sustained commensal dysbiosis in mice. PLoS ONE, 13.","DOI":"10.1101\/247866"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1007\/978-1-4939-9857-9_15","article-title":"Toxoplasma gondii: Bradyzoite Differentiation In Vitro and In Vivo","volume":"2071","author":"Mayoral","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1016\/j.vaccine.2010.11.002","article-title":"Evaluation of three recombinant multi-antigenic vaccines composed of surface and secretory antigens of Toxoplasma gondii in murine models of experimental toxoplasmosis","volume":"29","author":"Dziadek","year":"2011","journal-title":"Vaccine"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"55","DOI":"10.33073\/pjm-2015-007","article-title":"Recombinant MAG1 Protein of Toxoplasma gondii as a Diagnostic Antigen","volume":"64","author":"Gatkowska","year":"2015","journal-title":"Pol. J. Microbiol."},{"key":"ref_53","unstructured":"Sambrook, J., and Russell, D.W. (2001). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. [3rd ed.]."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1007\/978-1-4939-2269-7_22","article-title":"Drug affinity responsive target stability (DARTS) for small-molecule target identification","volume":"1263","author":"Pai","year":"2015","journal-title":"Methods Mol. Biol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/S0009-9120(97)00049-0","article-title":"Removal of endotoxin from recombinant protein preparations","volume":"30","author":"Liu","year":"1997","journal-title":"Clin. Biochem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3213\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:37:52Z","timestamp":1760135872000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/6\/3213"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,3,16]]},"references-count":55,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["ijms23063213"],"URL":"https:\/\/doi.org\/10.3390\/ijms23063213","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,3,16]]}}}