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For this purpose, both the loading capacity (LC) and Loading efficiency (LE) of the alginate NPs burdened by DT are evaluated. Moreover, the effects of different concentrations of sodium alginate and calcium chloride on the NPs physicochemical characteristics are surveyed in addition to other physical conditions such as homogenization time and rate. To do so, the NPs are characterised using particle size and distribution, zeta potential, scanning electron microscopy, encapsulation efficiency, in vitro release study and FT\u2010IR spectroscopy. Subsequently, the effects of homogenization time and rate on the NPs are assessed. At the meantime, the NPs LC and efficiency in several DT concentrations are estimated. The average size of the NPs was 400.7 and 276.6\u00a0nm for unloaded and DT loaded, respectively. According to the obtained results, the zeta potential of the blank and DT loaded NPs are estimated as \u221223.7\u00a0mV and \u221221.2\u00a0mV, respectively. Whereas, the LC and LE were &gt;80% and &gt;90%, in that order. Furthermore, 95% of the releasing DT loaded NPs occurs at 140\u00a0h in the sustained mode without any bursting release. It can be concluded that the features of NPs such as morphology and particle size are strongly depended on the calcium chloride, sodium alginate concentrations and physicochemical conditions in the NPs formation process. In addition, appropriate concentrations of the sodium alginate and calcium ions would lead to obtaining the desirable NPs formation associated with the advantageous LE, LC (over 80%) and sustained in vitro release profile. Ultimately, the proposed NPs can be employed in vaccine formulation for the targeted delivery, controlled and slow antigen release associated with the improved antigen stability.<\/jats:p>","DOI":"10.1049\/nbt2.12088","type":"journal-article","created":{"date-parts":[[2022,5,24]],"date-time":"2022-05-24T23:49:17Z","timestamp":1653436157000},"page":"199-209","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Investigating preparation and characterisation of diphtheria toxoid\u2010loaded on sodium alginate nanoparticles"],"prefix":"10.1049","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1965-4357","authenticated-orcid":false,"given":"Samira","family":"Aghamiri","sequence":"first","affiliation":[{"name":"Department of Biology Science and Research Branch Islamic Azad University  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mojtaba","family":"Noofeli","sequence":"additional","affiliation":[{"name":"Department of Research and Development Razi Vaccine and Serum Research Institute Agriculture Research Education and Extension Organization (AREEO)  Karaj Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Parvaneh","family":"Saffarian","sequence":"additional","affiliation":[{"name":"Department of Biology Science and Research Branch Islamic Azad University  Tehran Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zahra","family":"Salehi Najafabadi","sequence":"additional","affiliation":[{"name":"Department of Human Bacterial Vaccine Razi Vaccine and Serum Research Institute Agriculture Research Education and Extension Organization (AREEO)  Karaj Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hamid Reza","family":"Goudarzi","sequence":"additional","affiliation":[{"name":"Department of Research and Development Razi Vaccine and Serum Research Institute Agriculture Research Education and Extension Organization (AREEO)  Karaj Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"265","published-online":{"date-parts":[[2022,5,24]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1049\/iet\u2010nbt.2014.0062"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1049\/mnl.2019.0355"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2013.00013"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1007\/12_2011_150"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro1681"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2009.12.084"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.4172\/2157-7560.1000115"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1002\/path.1700290106"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/0264\u2010410x(93)90190\u20109"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.4103\/2230\u2010973x.82384"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.3233\/hab\u2010170324"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1049\/iet\u2010opt.2013.0080"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/12_2011_150"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1586\/erv.10.89"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/s0168\u20103659(00)00339\u20104"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/s1359\u20100286(02)00117\u20101"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1049\/iet\u2010nbt.2018.5248"},{"issue":"4","key":"e_1_2_10_19_1","first-page":"125","article-title":"Nanoparticles in vaccine development","volume":"1","author":"Hajizade A.","year":"2014","journal-title":"J. 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