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Quantitative analysis via SDS-PAGE densitometry revealed that the addition of 40% (v\/v) DMSO significantly enhanced the reaction yield compared to the aqueous control, while methanol resulted in a marked decrease in efficiency. These results were rationalized using Hansen solubility parameters, which indicated that the high dispersion cohesion (18.4 MPa1\/2) of DMSO promotes superior solvation of PEG chains, thereby minimizing competitive hydrolysis pathways. Furthermore, the inclusion of nonfunctionalized PEGs (4 and 8 kDa) provided a marginal increase in conjugation, likely due to the excluded volume effect. In contrast, the anionic surfactant SDS severely inhibited the reaction, a phenomenon attributed to electrostatic repulsion between the charged SDS-PEG-protein complexes. The nonionic surfactant polysorbate 20 exhibited a neutral effect on yield, suggesting its utility primarily as a protein stabilizer without compromising conjugation. These findings provide a comprehensive framework for the strategic selection of additives to modulate the physicochemical environment and maximize PEGylation efficiency.<\/jats:p>","DOI":"10.1021\/acsomega.6c06664","type":"journal-article","created":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T15:04:03Z","timestamp":1790089443000},"page":"59773-59783","source":"Crossref","is-referenced-by-count":0,"title":["Impact of Organic\nSolvents, Surfactants, and Nonfunctionalized\nPEGs on PEGylation Reaction Yield"],"prefix":"10.1021","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-8619-3286","authenticated-orcid":true,"given":"Raissa H. S.","family":"Florindo","sequence":"first","affiliation":[{"name":"Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-909"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea M.","family":"Salgado","sequence":"additional","affiliation":[{"name":"Universidade Federal do Rio de Janeiro , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-909"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5427-1928","authenticated-orcid":true,"given":"Vivian M.","family":"Saez","sequence":"additional","affiliation":[{"name":"Universidade Federal do Rio de Janeiro , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-909"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tito L.","family":"Moitinho-Alves","sequence":"additional","affiliation":[{"name":"Cidade Universita\u0301ria , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-909"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9174-582X","authenticated-orcid":true,"given":"Jose A.","family":"Ramon","sequence":"additional","affiliation":[{"name":"Universidade Federal do Rio de Janeiro , , , ,","place":["Rio de Janeiro, Rio de Janeiro, Brazil, 21941-909"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,22]]},"reference":[{"issue":"7","key":"2026100610481830100_cit1","doi-asserted-by":"publisher","first-page":"743","DOI":"10.2174\/1389200216666141208151524","article-title":"Protein engineering:\na new frontier for biological therapeutics","volume":"15","author":"Tobin","year":"2014","journal-title":"Curr. 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