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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:ali="http://www.niso.org/schemas/ali/1.0/" article-type="research-article" dtd-version="1.2" xml:lang="en"><front><journal-meta><journal-id journal-id-type="publisher-id">Current Pharmaceutical Biotechnology</journal-id><journal-title-group><journal-title xml:lang="en">Current Pharmaceutical Biotechnology</journal-title><trans-title-group xml:lang="ru"><trans-title>Current Pharmaceutical Biotechnology</trans-title></trans-title-group></journal-title-group><issn publication-format="print">1389-2010</issn><issn publication-format="electronic">1873-4316</issn><publisher><publisher-name xml:lang="en">Bentham Science</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="publisher-id">644758</article-id><article-id pub-id-type="doi">10.2174/1389201024666230519144615</article-id><article-categories><subj-group subj-group-type="toc-heading"><subject>Biotechnology</subject></subj-group><subj-group subj-group-type="article-type"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title xml:lang="en">Characterization and Immunogenicity of Recombinant A. flavus Uox Modified by Co/EDTA Carbon Dots</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Li</surname><given-names>Hai-Ling</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><name><surname>Gao</surname><given-names>Xiu-Feng</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Jing-Ji</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff2"/></contrib><contrib contrib-type="author"><name><surname>Wan</surname><given-names>Ming-Xia</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Guo-Qi</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff3"/></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Yong-Sheng</given-names></name><email>info@benthamscience.net</email><xref ref-type="aff" rid="aff4"/></contrib></contrib-group><aff id="aff1"><institution>West China School of Basic Medical Sciences &amp; Forensic Medicine, Sichuan University</institution></aff><aff id="aff2"><institution>College of Ecology and Environment,, Chengdu University of Technology</institution></aff><aff id="aff3"><institution>Department of Chemistry, School of Science, Xihua University</institution></aff><aff id="aff4"><institution>School of Chemical Engineering, Sichuan University</institution></aff><pub-date date-type="pub" iso-8601-date="2024-01-15" publication-format="electronic"><day>15</day><month>01</month><year>2024</year></pub-date><volume>25</volume><issue>2</issue><issue-title xml:lang="ru"/><fpage>230</fpage><lpage>246</lpage><history><date date-type="received" iso-8601-date="2025-01-07"><day>07</day><month>01</month><year>2025</year></date></history><permissions><copyright-statement xml:lang="en">Copyright ©; 2024, Bentham Science Publishers</copyright-statement><copyright-year>2024</copyright-year><copyright-holder xml:lang="en">Bentham Science Publishers</copyright-holder><ali:free_to_read xmlns:ali="http://www.niso.org/schemas/ali/1.0/"/></permissions><self-uri xlink:href="https://journals.eco-vector.com/1389-2010/article/view/644758">https://journals.eco-vector.com/1389-2010/article/view/644758</self-uri><abstract xml:lang="en"><p id="idm46041443605152">Background:Uricase (Uox) is a major drug in gout and a supplementary drug in cancer treatment. Because allergic reactions caused by Uox limit its clinical application,10% Co/EDTA was used to chemically modify Uox from A. flavus to reduce its immunogenicity.</p><p id="idm46041443609152">Methods:The immunogenicity of Uox and 10% Co/EDTA-Uox was examined by determining the antibody titer and concentration of IL-2, IL-6, IL-10, and TNF-β in quail and rat serum. Moreover, we examined the pharmacokinetics of 10% Co/EDTA-Uox in rats and acute toxicity in mice.</p><p id="idm46041443613120">Results:The concentration of UA decreased from 771.85 ± 180.99 to 299.47 ± 20.37 µmoL/L (p(&lt;0.01) in the hyperuricemia model of quails injected by 10% Co/EDTA-Uox. Two-way immuno- diffusion electrophoresis revealed that 10% Co/EDTA-Uox did not produce antibody, whereas the antibody titer against Uox was 1:16. The concentrations of four cytokines in the 10% Co/EDTA-Uox group were significantly lower than in Uox group (p &lt; 0.01); The titer of IgG and IgM against 10% Co/EDTA-Uox was significantly lower than that against Uox at different serum dilutions (p &lt; 0.0001). The pharmacokinetic data indicated that the half-life time of 10% Co/EDTA- Uox (69.315 h) was significantly longer than that of Uox (13.4 h) (p(&lt;0.01). The tissue section of the liver, heart, kidney, and spleen revealed no toxicity in Uox and 10% Co/EDTA- Uox groups.</p><p id="idm46041443618176">Conclusion:10% Co/EDTA-Uox possesses little immunogenicity, a long half-life time, and a highly efficient degradation of UA.</p></abstract><kwd-group xml:lang="en"><kwd>UA</kwd><kwd>hyperuricemia</kwd><kwd>Uox</kwd><kwd>Co/EDTA-Uox</kwd><kwd>cytotoxicity</kwd><kwd>immunogenicity.</kwd></kwd-group></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Li, Z. Phylogenetic articulation of uric acid evolution in mammals and how it informs a therapeutic uriase. Mol. Biol. Evol., 2021, 39(1), 1-8. doi: 10.1093/molbev/msab312</mixed-citation></ref><ref id="B2"><label>2.</label><mixed-citation>Perez-Ruiz, F.M.D. Gout. Rheum. Dis. Clin. North Am., 2019, 4(45), 583-591.</mixed-citation></ref><ref id="B3"><label>3.</label><mixed-citation>Dehlin, M.; Drivelegka, P.; Sigurdardottir, V.; Svärd, A.; Jacobsson, L.T.H. Incidence and prevalence of gout in Western Sweden. Arthritis Res. Ther., 2016, 18(1), 164-170. doi: 10.1186/s13075-016-1062-6 PMID: 27412614</mixed-citation></ref><ref id="B4"><label>4.</label><mixed-citation>Rai, S.K.; Aviña-Zubieta, J.A.; McCormick, N. The rising prevalence and incidence of gout in British Columbia, Canada: Population-based trends from 2000 to 2012. Semin. Arthritis Rheum., 2017, 46, 451-456.</mixed-citation></ref><ref id="B5"><label>5.</label><mixed-citation>Kim, J.W.; Kwak, S.G.; Lee, H.; Kim, S.K.; Choe, J.Y.; Park, S.H. Prevalence and incidence of gout in Korea: Data from the national health claims database 20072015. Rheumatol. Int., 2017, 37(9), 1499-1506. doi: 10.1007/s00296-017-3768-4 PMID: 28676911</mixed-citation></ref><ref id="B6"><label>6.</label><mixed-citation>Kuo, C.F.; Grainge, M.J.; Mallen, C.; Zhang, W.; Doherty, M. Rising burden of gout in the UK but continuing suboptimal management: a nationwide population study. Ann. Rheum. Dis., 2015, 74(4), 661-667. doi: 10.1136/annrheumdis-2013-204463 PMID: 24431399</mixed-citation></ref><ref id="B7"><label>7.</label><mixed-citation>Zobbe, K.; Prieto-Alhambra, D.; Cordtz, R.; Højgaard, P.; Hindrup, J.S.; Kristensen, L.E.; Dreyer, L. Secular trends in the incidence and prevalence of gout in Denmark from 1995 to 2015: a nationwide register-based study. Rheumatology (Oxford), 2019, 58(5), 836-839. doi: 10.1093/rheumatology/key390 PMID: 30590724</mixed-citation></ref><ref id="B8"><label>8.</label><mixed-citation>Chen-Xu, M.; Yokose, C.; Rai, S.K.; Pillinger, M.H.; Choi, H.K. Contemporary prevalence of gout and hyperuricemia in the United States and decadal trends: The National Health and Nutrition Examination Survey, 20072016. Arthritis Rheumatol., 2019, 71(6), 991-999. doi: 10.1002/art.40807 PMID: 30618180</mixed-citation></ref><ref id="B9"><label>9.</label><mixed-citation>Drivelegka, P.; Sigurdardottir, V.; Svärd, A.; Jacobsson, L.T.H.; Dehlin, M. Comorbidity in gout at the time of first diagnosis: sex differences that may have implications for dosing of urate lowering therapy. Arthritis Res. Ther., 2018, 20(1), 108-119. doi: 10.1186/s13075-018-1596-x PMID: 29855389</mixed-citation></ref><ref id="B10"><label>10.</label><mixed-citation>Vitart, V.; Rudan, I.; Hayward, C.; Gray, N.K.; Floyd, J.; Palmer, C.N.A.; Knott, S.A.; Kolcic, I.; Polasek, O.; Graessler, J.; Wilson, J.F.; Marinaki, A.; Riches, P.L.; Shu, X.; Janicijevic, B.; Smolej-Narancic, N.; Gorgoni, B.; Morgan, J.; Campbell, S.; Biloglav, Z.; Barac-Lauc, L.; Pericic, M.; Klaric, I.M.; Zgaga, L.; Skaric-Juric, T.; Wild, S.H.; Richardson, W.A.; Hohenstein, P.; Kimber, C.H.; Tenesa, A.; Donnelly, L.A.; Fairbanks, L.D.; Aringer, M.; McKeigue, P.M.; Ralston, S.H.; Morris, A.D.; Rudan, P.; Hastie, N.D.; Campbell, H.; Wright, A.F. SLC2A9 is a newly identified urate transporter influencing serum urate concentration, urate excretion and gout. Nat. Genet., 2008, 40(4), 437-442. doi: 10.1038/ng.106 PMID: 18327257</mixed-citation></ref><ref id="B11"><label>11.</label><mixed-citation>Enomoto, A.; Kimura, H.; Chairoungdua, A.; Shigeta, Y.; Jutabha, P.; Ho Cha, S.; Hosoyamada, M.; Takeda, M.; Sekine, T.; Igarashi, T.; Matsuo, H.; Kikuchi, Y.; Oda, T.; Ichida, K.; Hosoya, T.; Shimokata, K.; Niwa, T.; Kanai, Y.; Endou, H. Molecular identification of a renal urateanion exchanger that regulates blood urate levels. Nature, 2002, 417(6887), 447-452. doi: 10.1038/nature742 PMID: 12024214</mixed-citation></ref><ref id="B12"><label>12.</label><mixed-citation>Ekaratanawong, S.; Anzai, N.; Jutabha, P. Human organic anion transporter 4 is a renal apical organic anion/dicarboxylate exchanger in the proximal tubules. J. Pharmacol. Sci., 2004, 94, 297-304.</mixed-citation></ref><ref id="B13"><label>13.</label><mixed-citation>Eraly, S.A.; Vallon, V.; Rieg, T. Multiple organic anion transporters contribute to net renal excretion of uric acid. Physiol. Genomics, 2008, 33, 180-192. doi: 10.1152/physiolgenomics.00207.2007</mixed-citation></ref><ref id="B14"><label>14.</label><mixed-citation>Chhana, A.; Pool, B.; Wei, Y.; Choi, A.; Gao, R.; Munro, J.; Cornish, J.; Dalbeth, N. Human cartilage homogenates influence the crystallization of monosodium urate and inflammatory response to monosodium urate crystals: A potential link between osteoarthritis and gout. Arthritis Rheumatol., 2019, 71(12), 2090-2099. doi: 10.1002/art.41038 PMID: 31297987</mixed-citation></ref><ref id="B15"><label>15.</label><mixed-citation>Li, H.; Huo, J.; Sun, D.; Guo, Y.; Jiang, L.; Zhang, H.; Shi, X.; Zhao, Z.; Zhou, J.; Hu, C.; Zhang, C. Determination of PEGylation homogeneity of polyethylene glycol‐modified canine uricase. Electrophoresis, 2021, 42(6), 693-699. doi: 10.1002/elps.202000268 PMID: 33247595</mixed-citation></ref><ref id="B16"><label>16.</label><mixed-citation>Nyborg, A.C.; Ward, C.; Zacco, A.; Chacko, B.; Grinberg, L.; Geoghegan, J.C.; Bean, R.; Wendeler, M.; Bartnik, F.; OConnor, E.; Gruia, F.; Iyer, V.; Feng, H.; Roy, V.; Berge, M.; Miner, J.N.; Wilson, D.M.; Zhou, D.; Nicholson, S.; Wilker, C.; Wu, C.Y.; Wilson, S.; Jermutus, L.; Wu, H.; Owen, D.A.; Osbourn, J.; Coats, S.; Baca, M.A. Therapeutic uricase with reduced immunogenicity risk and improved development properties. PLoS One, 2016, 11(12), e0167935. doi: 10.1371/journal.pone.0167935</mixed-citation></ref><ref id="B17"><label>17.</label><mixed-citation>Bomalaski, M.J.S.; Goddard, D.H.; Grezlak, D.; Lopatin, M.A.; Holtsberg, F.W.; Ensor, C.M.; Clark, M.A. Clark, Phase I study of uricase formulated with polyethylene glycol (Uricase-PEG 20), Abstract 287.American College of Rheumatology Annual Scientific Meeting; New Orleans, LA, 2002, p. 25-29.</mixed-citation></ref><ref id="B18"><label>18.</label><mixed-citation>Pooja, N. JagadeeshBabu, P.E. Studies on the site-specifific PEGylation induced interferences instigated in uricase quantifification using the bradford method. Int. J. Pept. Res. The., 2016, 16, 1-9. doi: 10.1007/s10989-016-9518-8</mixed-citation></ref><ref id="B19"><label>19.</label><mixed-citation>Xiaopei, Zh.; Duo, X.; Xin, J. Nanocapsules of therapeutic proteins with enhanced stability and long blood circulation for hyperuricemia management. J. Control. Release, 2017, 2017(255), 54-61. doi: 10.1016/j.jconrel.2017.03.019</mixed-citation></ref><ref id="B20"><label>20.</label><mixed-citation>Chen, J.W.T. DNA shuffling of uricase gene leads to a more "human like" chimeric uricase with increased uricolytic activity. Int. J. Biol. Macromol., 2016, 82, 522-529.</mixed-citation></ref><ref id="B21"><label>21.</label><mixed-citation>Nelapati, A.K.; Das, B.K.; Ponnan, E.J.B.; Chakraborty, D. In-silico epitope identification and design of Uricase mutein with reduced immunogenicity. Process Biochem., 2020, 92(92), 288-302. doi: 10.1016/j.procbio.2020.01.022</mixed-citation></ref><ref id="B22"><label>22.</label><mixed-citation>Sands, E.; Kivitz, A.; DeHaan, W.; Leung, S.S.; Johnston, L.; Kishimoto, T.K. Tolerogenic nanoparticles mitigate the formation of anti-drug antibodies against pegylated uricase in patients with hyperuricemia. Nat. Commun., 2022, 13(1), 272-286. doi: 10.1038/s41467-021-27945-7 PMID: 35022448</mixed-citation></ref><ref id="B23"><label>23.</label><mixed-citation>Schlesinger, N.; Padnick-Silver, L.; LaMoreaux, B. Enchancing the response rate to recombianat uricases in patients with gout. BioDrugs, 2022, 36(2), 95-103. doi: 10.1007/s40259-022-00517-x</mixed-citation></ref><ref id="B24"><label>24.</label><mixed-citation>Wu, J.; Chen, G.; Jia, Y.; Ji, C.; Wang, Y.; Zhou, Y.; Leblanc, R.M.; Peng, Z. Carbon dot composites for bioapplications: A review. J. Mater. Chem. B Mater. Biol. Med., 2022, 10(6), 843-869. doi: 10.1039/D1TB02446A PMID: 35060567</mixed-citation></ref><ref id="B25"><label>25.</label><mixed-citation>Zhang, G.Q.; Li, Y.; Liu, W.P.; Gao, X.F. A fluorimetric and colorimetric dual-signal sensor for hydrogen peroxide and glucose based on the intrinsic peroxidase-like activity of cobalt and nitrogen co-doped carbon dots and inner filter effect. Anal. Methods, 2021, 13(28), 3196-3204. doi: 10.1039/D1AY00781E PMID: 34184019</mixed-citation></ref><ref id="B26"><label>26.</label><mixed-citation>Rao, C.; Khan, S.; Verma, N.C.; Nandi, C.K. Labelling Proteins with carbon nanodots. ChemBioChem, 2017, 18(24), 2385-2389. doi: 10.1002/cbic.201700440 PMID: 28985453</mixed-citation></ref><ref id="B27"><label>27.</label><mixed-citation>Silva, A.C.A.; Freschi, A.P.P.; Rodrigues, C.M.; Matias, B.F.; Maia, L.P.; Goulart, L.R.; Dantas, N.O. Biological analysis and imaging applications of CdSe/CdSxSe1−x/CdS coreshell magic-sized quantum dot. Nanomedicine, 2016, 12(5), 1421-1430. doi: 10.1016/j.nano.2016.01.001</mixed-citation></ref><ref id="B28"><label>28.</label><mixed-citation>Kokorina, A.A.; Bakal, A.A.; Shpuntova, D.V.; Kostritskiy, A.Y.; Beloglazova, N.V.; De Saeger, S.; Sukhorukov, G.B.; Sapelkin, A.V.; Goryacheva, I.Y. Gel electrophoresis separation and origins of light emission in fluorophores prepared from citric acid and ethylenediamine. Sci. Rep., 2019, 9(1), 14665. doi: 10.1038/s41598-019-50922-6 PMID: 31605021</mixed-citation></ref><ref id="B29"><label>29.</label><mixed-citation>Kuznetsova, V.A.; Visheratina, A.K.; Ryan, A.; Martynenko, I.V.; Loudon, A.; Maguire, C.M.; Purcell-Milton, F.; Orlova, A.O.; Baranov, A.V.; Fedorov, A.V.; Prina-Mello, A.; Volkov, Y. GunKo, Y.K. Enantioselective cytotoxicity of ZnS:Mn quantum dots in A549 cells. Chirality, 2017, 29(8), 403-408. doi: 10.1002/chir.22713 PMID: 28608629</mixed-citation></ref><ref id="B30"><label>30.</label><mixed-citation>Yan, M.; Ge, J.; Liu, Z.; Ouyang, P. Encapsulation of single enzyme in nanogel with enhanced biocatalytic activity and stability. J. Am. Chem. Soc., 2006, 128(34), 11008-11009. doi: 10.1021/ja064126t PMID: 16925402</mixed-citation></ref><ref id="B31"><label>31.</label><mixed-citation>Yunli, Zh.; Mi, Zh.; Dan, H. Uricase alkaline enzymosomes with enhanced stabilities and anti hyperuricemia effects induced by favorable microEnvironmental changes; Scientific Repots, 2016. doi: 10.1038/srep20136</mixed-citation></ref><ref id="B32"><label>32.</label><mixed-citation>Zhang, P.; Sun, F.; Tsao, C.; Liu, S.; Jain, P.; Sinclair, A.; Hung, H.C.; Bai, T.; Wu, K.; Jiang, S. Zwitterionic gel encapsulation promotes protein stability, enhances pharmacokinetics, and reduces immunogenicity. Proc. Natl. Acad. Sci. USA, 2015, 112(39), 12046-12051. doi: 10.1073/pnas.1512465112 PMID: 26371311</mixed-citation></ref><ref id="B33"><label>33.</label><mixed-citation>Kim, S.; Kim, M.; Jung, S.; Kwon, K.; Park, J.; Kim, S.; Kwon, I.; Tae, G. Co-delivery of therapeutic protein and catalase-mimic nanoparticle using a biocompatible nanocarrier for enhanced therapeutic effect. J. Control. Release, 2019, 309, 181-189. doi: 10.1016/j.jconrel.2019.07.038 PMID: 31356840</mixed-citation></ref><ref id="B34"><label>34.</label><mixed-citation>Ming, J.; Zhu, T.; Li, J.; Ye, Z.; Shi, C.; Guo, Z.; Wang, J.; Chen, X.; Zheng, N. A Novel cascade nanoreactor integrating Two-Dimensional Pd-Ru nanozyme, uricase and red blood cell membrane for highly efficient hyperuricemia treatment. Nano-micro. Small, 2021, 17(46), 2103645. doi: 10.1002/smll.202103645</mixed-citation></ref><ref id="B35"><label>35.</label><mixed-citation>Zhang, P.; Jain, P.; Tsao, C.; Yuan, Z.; Li, W.; Li, B.; Wu, K.; Hung, H-C.; Lin, X.; Jiang, S. Polypeptides with high zwitterion density for safe and effective therapeutics. Angew. Chem., 2018, 130(26), 7869-7873. doi: 10.1002/ange.201802452</mixed-citation></ref><ref id="B36"><label>36.</label><mixed-citation>da Silva Freitas, D.; Spencer, P.J.; Vassão, R.C.; Abrahão-Neto, J. Biochemical and biopharmaceutical properties of PEGylated uricase. Int. J. Pharm., 2010, 387(1-2), 215-222. doi: 10.1016/j.ijpharm.2009.11.034 PMID: 19969053</mixed-citation></ref><ref id="B37"><label>37.</label><mixed-citation>Jun-ichi, T.; Katsumi, H.; Etsuko, K.; Maki, N.; Itary, Y. Studies on antigenicity of the polyethylene glycol (PEG)-modified uricase. Int. J. Immunopharmacol., 1985, 7(5), 725-730. doi: 10.1016/0192-0561(85)90158-4 PMID: 2412977</mixed-citation></ref></ref-list></back></article>
