{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T09:29:49Z","timestamp":1783330189473,"version":"3.54.6"},"reference-count":63,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T00:00:00Z","timestamp":1733788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Introduction<\/jats:title><jats:p>Immunogenicity continues to be a challenge for development and clinical utility of monoclonal antibodies, and there are gaps in our current ability to prevent anti-drug antibody development in a safe and antigen-specific manner.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>To mitigate immunogenicity of monoclonal antibodies administered subcutaneously, O-phospho-L-serine (OPLS)\u2014the head group of the tolerance-inducing phospholipid, phosphatidylserine\u2014was investigated as an immunoregulatory adjuvant.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Formulations of adalimumab, trastuzumab or rituximab with OPLS showed reduction in relative immunogenicity in mice compared to vehicle formulations, indicated by reduced anti-drug antibody development and significant reductions in CD138+ plasma cell differentiation in bone marrow. Titer development toward recombinant human hyaluronidase, a dispersion enhancer that was co-formulated with monoclonal antibodies, was similarly reduced. Subcutaneous administration of adalimumab with OPLS resulted in a two-fold increase in expression of type 1 regulatory (Tr1) T cell subset in the spleen. This is consistent with in vitro studies where co-culturing of dendritic cells primed with ovalbumin in the presence and absence of OPLS and antigen specific T-cells induced expression of Tr1 phenotype on live CD4+ T cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>This adjuvant does not impact immune competence of non-human primates and mice, and repeated administration of the adjuvant does not show renal or hepatic toxicity. Formulation of monoclonal antibodies with the immunoregulatory adjuvant, OPLS, was found to be safe and effective at mitigating immunogenicity.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2024.1496169","type":"journal-article","created":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T05:11:00Z","timestamp":1733807460000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Immune regulatory adjuvant approach to mitigate subcutaneous immunogenicity of monoclonal antibodies"],"prefix":"10.3389","volume":"15","author":[{"given":"Nicole L.","family":"Jarvi","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manali","family":"Patel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Krithika A.","family":"Shetty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nhan H.","family":"Nguyen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brooke F.","family":"Grasperge","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Donald E.","family":"Mager","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert M.","family":"Straubinger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sathy V.","family":"Balu-Iyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,12,10]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2018.01.007","article-title":"Challenges and opportunities for the subcutaneous delivery of therapeutic proteins","volume":"107","author":"Turner","year":"2018","journal-title":"J Pharm Sci"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-020-00465-4","article-title":"Immunogenicity challenges associated with subcutaneous delivery of therapeutic proteins","volume":"35","author":"Jarvi","year":"2021","journal-title":"BioDrugs"},{"key":"B3","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1177\/0091270012436560","article-title":"Comparison of subcutaneous and intravenous administration of trastuzumab: a phase I\/Ib trial in healthy male volunteers and patients with HER2-positive breast cancer","volume":"53","author":"Wynne","year":"2013","journal-title":"J Clin Pharmacol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2012.110","article-title":"Bortezomib in the rapid reduction of high sustained antibody titers in disorders treated with therapeutic protein: lessons learned from Pompe disease","volume":"15","author":"Banugaria","year":"2013","journal-title":"Genet Med"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2011.4","article-title":"Successful immune tolerance induction to enzyme replacement therapy in CRIM-negative infantile Pompe disease","volume":"14","author":"Messinger","year":"2012","journal-title":"Genet Med"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1055\/s-0038-1649664","article-title":"A prospective, randomized trial of prednisone and cyclophosphamide in the treatment of patients with factor VIII autoantibodies","volume":"70","author":"Green","year":"1993","journal-title":"Thromb Haemost"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1002\/1529-0131(199809)41:9<1552::AID-ART5>3.0.CO;2-W","article-title":"Therapeutic efficacy of multiple intravenous infusions of anti-tumor necrosis factor alpha monoclonal antibody combined with low-dose weekly methotrexate in rheumatoid arthritis","volume":"41","author":"Maini","year":"1998","journal-title":"Arthritis Rheum"},{"key":"B8","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1080\/08820139.2020.1738456","article-title":"Biological function and immunotherapy utilizing phosphatidylserine-based nanoparticles","volume":"49","author":"Glassman","year":"2020","journal-title":"Immunol Invest"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2018.03.027","article-title":"Phosphatidylserine is not just a cleanup crew but also a well-meaning teacher","volume":"107","author":"Glassman","year":"2018","journal-title":"J Pharm Sci"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.C112.396325","article-title":"Exposure to factor VIII protein in the presence of phosphatidylserine induces hypo-responsiveness toward factor VIII challenge in hemophilia A mice","volume":"288","author":"Gaitonde","year":"2013","journal-title":"J Biol Chem"},{"key":"B11","doi-asserted-by":"publisher","first-page":"17853","DOI":"10.1038\/s41598-021-97333-0","article-title":"Rational design of a nanoparticle platform for oral prophylactic immunotherapy to prevent immunogenicity of therapeutic proteins","volume":"11","author":"Nguyen","year":"2021","journal-title":"Sci Rep"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.1016\/j.xphs.2016.06.018","article-title":"Phosphatidylserine converts immunogenic recombinant human acid alpha-glucosidase to a tolerogenic form in a mouse model of pompe disease","volume":"105","author":"Schneider","year":"2016","journal-title":"J Pharm Sci"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpharm.2018.07.018","article-title":"Subcutaneous administration of Lyso-phosphatidylserine nanoparticles induces immunological tolerance towards Factor VIII in a Hemophilia A mouse model","volume":"548","author":"Glassman","year":"2018","journal-title":"Int J Pharm"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1038\/mi.2012.100","article-title":"TIM-4, expressed by medullary macrophages, regulates respiratory tolerance by mediating phagocytosis of antigen-specific T cells","volume":"6","author":"Albacker","year":"2013","journal-title":"Mucosal Immunol"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1001360","article-title":"TIM-4, a receptor for phosphatidylserine, controls adaptive immunity by regulating the removal of antigen-specific T cells","volume":"185","author":"Albacker","year":"2010","journal-title":"J Immunol (Baltimore Md: 1950)"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1038\/nature06307","article-title":"Identification of Tim4 as a phosphatidylserine receptor","volume":"450","author":"Miyanishi","year":"2007","journal-title":"Nature"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2007.11.011","article-title":"TIM-1 and TIM-4 glycoproteins bind phosphatidylserine and mediate uptake of apoptotic cells","volume":"27","author":"Kobayashi","year":"2007","journal-title":"Immunity"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejps.2014.10.010","article-title":"Intravenous administration of Factor VIII-O-Phospho-L-Serine (OPLS) complex reduces immunogenicity and preserves pharmacokinetics of the therapeutic protein","volume":"66","author":"Gaitonde","year":"2015","journal-title":"Eur J Pharm Sci"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1208\/aapsj0902028","article-title":"O-phospho-L-serine, multi-functional excipient for B domain deleted recombinant factor VIII","volume":"9","author":"Miclea","year":"2007","journal-title":"AAPS J"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M500163200","article-title":"Lower inhibitor development in hemophilia A mice following administration of recombinant factor VIII-O-phospho-L-serine complex","volume":"280","author":"Purohit","year":"2005","journal-title":"J Biol Chem"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1002\/jps.24173","article-title":"O-Phospho-l-Serine mediates Hyporesponsiveness toward FVIII in Hemophilia A-Murine Model by Inducing Tolerogenic Properties in Dendritic Cells","volume":"103","author":"Fathallah","year":"2014","journal-title":"J Pharm Sci"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.638444","article-title":"Efficacy and safety of adalimumab biosimilars: current critical clinical data in rheumatoid arthritis","volume":"12","author":"Lu","year":"2021","journal-title":"Front Immunol"},{"key":"B23","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1080\/10717544.2018.1551442","article-title":"ENHANZE(\u00ae) drug delivery technology: a novel approach to subcutaneous administration using recombinant human hyaluronidase PH20","volume":"26","author":"Locke","year":"2019","journal-title":"Drug delivery"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1038\/bjc.2013.371","article-title":"Subcutaneous administration of rituximab (MabThera) and trastuzumab (Herceptin) using hyaluronidase","volume":"109","author":"Shpilberg","year":"2013","journal-title":"Br J Cancer"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.jtho.2024.05.005","article-title":"Brief report: updated data from IMscin001 part 2, a randomized phase III study of subcutaneous versus intravenous atezolizumab in patients with locally advanced\/metastatic NSCLC","volume":"19","author":"Burotto","year":"2024","journal-title":"J Thorac Oncol"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1002\/cpt.v115.3","article-title":"Model-based dose selection of subcutaneous nivolumab in patients with advanced solid tumors","volume":"115","author":"Zhao","year":"2024","journal-title":"Clin Pharmacol Ther"},{"key":"B27","doi-asserted-by":"publisher","first-page":"3597","DOI":"10.1212\/WNL.0000000000205244","article-title":"OCARINA II, phase III study: results of subcutaneous ocrelizumab administration in patients with multiple sclerosis (S31.006)","volume":"102","author":"Newsome","year":"2024","journal-title":"Neurology"},{"key":"B28","doi-asserted-by":"publisher","first-page":"6334","DOI":"10.1212\/WNL.0000000000206401","article-title":"Long-term safety, tolerability, and efficacy of subcutaneous efgartigimod PH20 in participants with generalized myasthenia gravis: interim results of the ADAPT-SC+ Study (P10-11.008)","volume":"102","author":"Howard","year":"2024","journal-title":"Neurology"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1208\/s12248-015-9782-0","article-title":"Clinical immunogenicity of rHuPH20, a hyaluronidase enabling subcutaneous drug administration","volume":"17","author":"Rosengren","year":"2015","journal-title":"AAPS J"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-018-0260-y","article-title":"Recombinant human PH20: baseline analysis of the reactive antibody prevalence in the general population using healthy subjects","volume":"32","author":"Rosengren","year":"2018","journal-title":"BioDrugs"},{"key":"B31","unstructured":"South San Francisco, CA\n          Genentech, Inc\n          Herceptin Hylecta\n          \n          2019"},{"key":"B32","unstructured":"South San Francisco, CA\n          Genentech, Inc\n          Rituxan Hycela\n          \n          2020"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1016\/0272-0590(84)90198-2","article-title":"Naphthalene toxicity in CD-1 mice: general toxicology and immunotoxicology","volume":"4","author":"Shopp","year":"1984","journal-title":"Fundam Appl Toxicol"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1007\/s40259-014-0103-4","article-title":"Comparative nonclinical assessments of the proposed biosimilar PF-05280014 and trastuzumab (Herceptin(\u00ae))","volume":"28","author":"Hurst","year":"2014","journal-title":"BioDrugs"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-014-1416-1","article-title":"Interspecies pharmacokinetic modeling of subcutaneous absorption of rituximab in mice and rats","volume":"31","author":"Kagan","year":"2014","journal-title":"Pharm Res"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)70226-3","article-title":"Phosphorus assay in column chromatography","volume":"234","author":"Bartlett","year":"1959","journal-title":"J Biol Chem"},{"key":"B37","doi-asserted-by":"publisher","first-page":"35","DOI":"10.1016\/S0022-1759(98)00170-7","article-title":"A statistically defined endpoint titer determination method for immunoassays","volume":"221","author":"Frey","year":"1998","journal-title":"J Immunol Methods"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-61779-012-6_16","article-title":"In vitro immunogenicity risk assessment of therapeutic proteins in preclinical setting","volume":"716","author":"Gaitonde","year":"2011","journal-title":"Methods Mol Biol"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-60761-401-2_11","article-title":"The T-dependent antibody response to keyhole limpet hemocyanin in rodents","volume":"598","author":"Plitnick","year":"2010","journal-title":"Methods Mol Biol"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1080\/15476910500362838","article-title":"T-cell-dependent antibody response: assay development in cynomolgus monkeys","volume":"2","author":"Piccotti","year":"2005","journal-title":"J Immunotoxicol"},{"key":"B41","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/106.4.549","article-title":"Advantage of using both anionic and zwitterionic phospholipid antigens for the detection of antiphospholipid antibodies","volume":"106","author":"Laroche","year":"1996","journal-title":"Am J Clin Pathol"},{"key":"B42","doi-asserted-by":"publisher","first-page":"7.14.1","DOI":"10.1002\/0471142735.2015.108.issue-1","article-title":"ELISPOT assay for measurement of antigen-specific and polyclonal antibody responses","volume":"108","author":"Lycke","year":"2015","journal-title":"Curr Protoc Immunol"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-019-0244-2","article-title":"B cell memory: building two walls of protection against pathogens","volume":"20","author":"Akkaya","year":"2020","journal-title":"Nat Rev Immunol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0132479","article-title":"Differences in CD44 surface expression levels and function discriminates IL-17 and IFN-\u03b3 Producing helper T cells","volume":"10","author":"Schumann","year":"2015","journal-title":"PloS One"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1038\/nm.3179","article-title":"Coexpression of CD49b and LAG-3 identifies human and mouse T regulatory type 1 cells","volume":"19","author":"Gagliani","year":"2013","journal-title":"Nat Med"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1016\/j.imlet.2003.11.004","article-title":"Induction of regulatory dendritic cells by dexamethasone and 1\u03b1,25-Dihydroxyvitamin D3","volume":"91","author":"Pedersen","year":"2004","journal-title":"Immunol Lett"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1101398","article-title":"Inducible CD4+LAP+Foxp3- regulatory T cells suppress allergic inflammation","volume":"187","author":"Duan","year":"2011","journal-title":"J Immunol"},{"key":"B48","first-page":"46","article-title":"Serum clinical chemistry and hematology reference values in outbred stocks of albino mice from three commonly used vendors and two inbred strains of albino mice","volume":"42","author":"Serfilippi","year":"2003","journal-title":"Contemp Top Lab Anim Sci"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1182\/asheducation-2005.1.462","article-title":"Thrombosis and the antiphospholipid syndrome","volume":"2005","author":"Ortel","year":"2005","journal-title":"Hematology"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1080\/17425247.2021.1981286","article-title":"Safety of recombinant human hyaluronidase PH20 for subcutaneous drug delivery","volume":"18","author":"Knowles","year":"2021","journal-title":"Expert Opin Drug Delivery"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1517\/17425247.4.4.427","article-title":"Recombinant human hyaluronidase (rHuPH20): an enabling platform for subcutaneous drug and fluid administration","volume":"4","author":"Frost","year":"2007","journal-title":"Expert Opin Drug Delivery"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1021\/bi00088a009","article-title":"Specific membrane binding of factor VIII is mediated by O-phospho-L-serine, a moiety of phosphatidylserine","volume":"32","author":"Gilbert","year":"1993","journal-title":"Biochemistry"},{"key":"B53","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1002\/(SICI)1520-6017(200003)89:3<297::AID-JPS2>3.0.CO;2-P","article-title":"Lymphatic transport of proteins after subcutaneous administration","volume":"89","author":"Porter","year":"2000","journal-title":"J Pharm Sci"},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2019.00965","article-title":"Survival of long-lived plasma cells (LLPC): piecing together the puzzle","volume":"10","author":"Lightman","year":"2019","journal-title":"Front Immunol"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2015.01.017","article-title":"Differentiation and maintenance of long-lived plasma cells","volume":"33","author":"Kometani","year":"2015","journal-title":"Curr Opin Immunol"},{"key":"B56","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.180.1.361","article-title":"Maintenance of long-lived plasma cells and serological memory despite mature and memory B cell depletion during CD20 immunotherapy in mice","volume":"180","author":"DiLillo","year":"2008","journal-title":"J Immunol"},{"key":"B57","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymgme.2012.01.019","article-title":"Persistence of high sustained antibodies to enzyme replacement therapy despite extensive immunomodulatory therapy in an infant with Pompe disease: need for agents to target antibody-secreting plasma cells","volume":"105","author":"Banugaria","year":"2012","journal-title":"Mol Genet Metab"},{"key":"B58","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2020.02.013","article-title":"PD-1: A driver or passenger of T cell exhaustion","volume":"77","author":"He","year":"2020","journal-title":"Mol Cell"},{"key":"B59","doi-asserted-by":"publisher","DOI":"10.14348\/molcells.2022.2056","article-title":"Current understanding of cytotoxic T lymphocyte antigen-4 (CTLA-4) signaling in T-cell biology and disease therapy","volume":"45","author":"Kim","year":"2022","journal-title":"Mol Cells"},{"key":"B60","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.25.21.9543-9553.2005","article-title":"CTLA-4 and PD-1 receptors inhibit T-cell activation by distinct mechanisms","volume":"25","author":"Parry","year":"2005","journal-title":"Mol Cell Biol"},{"key":"B61","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2008-02-138073","article-title":"Activation of natural regulatory T cells by IgG Fc-derived peptide \"Tregitopes","volume":"112","author":"De Groot","year":"2008","journal-title":"Blood"},{"key":"B62","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijpharm.2015.05.069","article-title":"Tolerability of hypertonic injectables","volume":"490","author":"Wang","year":"2015","journal-title":"Int J pharmaceutics"},{"key":"B63","unstructured":"Silver Spring, MD\n          FDA\n          Guidance for Industry: Estimating the Maximum Safe Starting Dose in Initial Clinical Trials for Therapeutics in Adult Healthy Volunteers\n          \n          2005"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1496169\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,12,10]],"date-time":"2024-12-10T05:11:07Z","timestamp":1733807467000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2024.1496169\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,10]]},"references-count":63,"alternative-id":["10.3389\/fimmu.2024.1496169"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2024.1496169","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,10]]},"article-number":"1496169"}}