{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,31]],"date-time":"2026-07-31T15:29:05Z","timestamp":1785511745590,"version":"3.56.0"},"reference-count":32,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T00:00:00Z","timestamp":1740096000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["1P01CA240307-01"],"award-info":[{"award-number":["1P01CA240307-01"]}]},{"name":"NIH","award":["P30CA013148"],"award-info":[{"award-number":["P30CA013148"]}]},{"name":"NEXT program","award":["1P01CA240307-01"],"award-info":[{"award-number":["1P01CA240307-01"]}]},{"name":"NEXT program","award":["P30CA013148"],"award-info":[{"award-number":["P30CA013148"]}]},{"name":"O\u2019Neal Comprehensive Cancer Center at UAB","award":["1P01CA240307-01"],"award-info":[{"award-number":["1P01CA240307-01"]}]},{"name":"O\u2019Neal Comprehensive Cancer Center at UAB","award":["P30CA013148"],"award-info":[{"award-number":["P30CA013148"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Objectives: Annexin-A1 is a 37 kDa phospholipid-binding protein which is concentrated in a truncated 34 kDa form (AnnA1) in caveolae on the tumor vascular endothelial cell surface with expression in many tumor types. PRISM developed the monoclonal mouse antibody mAnnA1 against AnnA1 for evaluation of AnnA1 as a potential target for imaging and therapy in oncology. mAnnA1 was humanized to make hAnnA1 for translation to clinical studies. Both PRISM-produced mAnnA1 and cGMP contractor-produced hAnnA1 were investigated using noninvasive PET\/CT imaging, and dosimetry was evaluated to enable clinical translation of this strategy and to investigate in vivo behavior of hAnnA1. Methods: Antibodies mAnnA1 and hAnnA1 (PRISM \u201chAnnA1-P\u201d or contractor generated \u201chAnnA1-C\u201d) were conjugated with the chelator deferoxamine and evaluated for immunoreactivity with ELISA. Conjugated antibodies were radiolabeled with zirconium-89. Na\u00efve mice, rats, and non-human primates (NHP) were injected with [89Zr]mAnnA1 or [89Zr]hAnnA1 and imaged with PET\/CT up to 10 days post injection. After imaging, mice and rats were euthanized and organs were collected, weighed, and radioactivity was quantified using a gamma counter. Dosimetry in mice and NHPs were calculated using OLINDA. Results: [89Zr]mAnnA1 showed similar biodistribution to other antibodies with slow clearance through the liver. Transition to [89Zr]hAnnA1-C during the dosimetry studies revealed substantial uptake in the spleen (130 \u00b1 48% ID\/g at day 5 post injection in female BALB\/c), which was not observed with [89Zr]mAnnA1 (5.6 \u00b1 1.7% ID\/g at day 7 PI). Further studies in multiple strains of mice showed variable elevated splenic uptake of [89Zr]hAnnA1-C across mouse strains, with the highest uptake observed in female BALB\/c mice (118.4 \u00b1 23.1% ID\/g) and the lowest uptake observed in male CD1 mice (34.7 \u00b1 10.2% ID\/g). Additionally, splenic uptake of hAnnA1-C was observed in Fischer rats (2.8 \u00b1 0.6% ID\/organ) and NHPs (1.6 \u00b1 0.6% ID\/organ), although at lower levels than what was observed in BALB\/c mice (8.8 \u00b1 1.8% ID\/organ). Dosimetry results showed similar values between estimates based on mouse and NHP data, with the largest difference seen in the spleen (5.2 vs. 2.6 mSv\/MBq in females respectively). Sequencing of hAnnA1-C revealed a frameshift mutation in the antibody sequence introduced during cGMP manufacture. Restoration of the antibody sequence by PRISM returned the antibody distribution into alignment with mAnnA1. Conclusions: An aberration introduced during cGMP production of hAnnA1-C resulted in increased splenic uptake and alteration of the biodistribution in mice. PET imaging enabled quantitative detection of the immunogenic behavior of hAnnA1, which led to detection of the sequence error. Restoration of the sequence resulted in an antibody which was non-immunogenic to mice.<\/jats:p>","DOI":"10.3390\/ph18030295","type":"journal-article","created":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T06:39:07Z","timestamp":1740119947000},"page":"295","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["PET Imaging Expedites Detection of Aberration in the Humanization of an Annexin A1 Targeting Antibody"],"prefix":"10.3390","volume":"18","author":[{"given":"Hailey A.","family":"Houson","sequence":"first","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian D.","family":"Wright","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Solana R.","family":"Fernandez","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tim","family":"Buss","sequence":"additional","affiliation":[{"name":"Proteogenomics Research Institute for Systems Medicine (PRISM), La Jolla, CA 92037, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sharon L.","family":"White","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brittany","family":"Cederstrom","sequence":"additional","affiliation":[{"name":"Proteogenomics Research Institute for Systems Medicine (PRISM), La Jolla, CA 92037, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James M.","family":"Omweri","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan E.","family":"McConathy","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8211-9539","authenticated-orcid":false,"given":"Jan E.","family":"Schnitzer","sequence":"additional","affiliation":[{"name":"Proteogenomics Research Institute for Systems Medicine (PRISM), La Jolla, CA 92037, USA"},{"name":"Institute of Engineering in Medicine, University of California San Diego, La Jolla, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0090-0191","authenticated-orcid":false,"given":"Suzanne E.","family":"Lapi","sequence":"additional","affiliation":[{"name":"Department of Radiology, University of Alabama at Birmingham, Birmingham, AL 35294, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1016\/j.addr.2020.06.013","article-title":"Targeting drug delivery in the vascular system: Focus on endothelium","volume":"157","author":"Glassman","year":"2020","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.jconrel.2017.09.016","article-title":"Overcoming key biological barriers to cancer drug delivery and efficacy","volume":"267","author":"Kim","year":"2017","journal-title":"J. Control. Release"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1062","DOI":"10.1038\/nm.3623","article-title":"In vivo proteomic imaging analysis of caveolae reveals pumping system to penetrate solid tumors","volume":"20","author":"Oh","year":"2014","journal-title":"Nat. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1038\/nri2470","article-title":"Annexin A1 and glucocorticoids as effectors of the resolution of inflammation","volume":"9","author":"Perretti","year":"2009","journal-title":"Nat. Rev. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1007\/s10585-011-9380-1","article-title":"Identification of annexin A1 as a proinvasive and prognostic factor for lung adenocarcinoma","volume":"28","author":"Liu","year":"2011","journal-title":"Clin. Exp. Metastasis"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1466","DOI":"10.3748\/wjg.v10.i10.1466","article-title":"Overexpression of annexin 1 in pancreatic cancer and its clinical significance","volume":"10","author":"Bai","year":"2004","journal-title":"World J. Gastroenterol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1827","DOI":"10.1007\/s12032-011-0013-y","article-title":"Proteome analysis of the effects of sorafenib on human hepatocellular carcinoma cell line HepG2","volume":"29","author":"Suo","year":"2012","journal-title":"Med. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/sj.bjc.6604128","article-title":"Characterisation and protein expression profiling of annexins in colorectal cancer","volume":"98","author":"Duncan","year":"2008","journal-title":"Br. J. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kadam, A.H., Kandasamy, K., Buss, T., Cederstrom, B., Yang, C., Narayanapillai, S., Rodriguez, J., Levin, M.D., Koziol, J., and Olenyuk, B. (2022). Targeting caveolae to pump bispecific antibody to TGF-\u03b2 into diseased lungs enables ultra-low dose therapeutic efficacy. PLoS ONE, 17.","DOI":"10.1101\/2022.09.12.507679"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.nucmedbio.2012.08.004","article-title":"PET imaging with 89Zr: From radiochemistry to the clinic","volume":"40","author":"Deri","year":"2013","journal-title":"Nucl. Med. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1007\/s11307-016-0951-z","article-title":"[89Zr]Trastuzumab: Evaluation of Radiation Dosimetry, Safety, and Optimal Imaging Parameters in Women with HER2-Positive Breast Cancer","volume":"18","author":"Laforest","year":"2016","journal-title":"Mol. Imaging Biol."},{"key":"ref_12","first-page":"164S","article-title":"The promise of immuno-PET in radioimmunotherapy","volume":"46","author":"Verel","year":"2005","journal-title":"J. Nucl. Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Lugat, A., Bailly, C., Cherel, M., Rousseau, C., Kraeber-Bodere, F., Bodet-Milin, C., and Bourgeois, M. (2022). Immuno-PET: Design options and clinical proof-of-concept. Front. Med., 9.","DOI":"10.3389\/fmed.2022.1026083"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Yoon, J.K., Park, B.N., Ryu, E.K., An, Y.S., and Lee, S.J. (2020). Current Perspectives on 89Zr-PET Imaging. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21124309"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"574","DOI":"10.1177\/01926233221088283","article-title":"Organ Weights in Relation to Age and Sex in Cynomolgus Monkeys (Macaca fascicularis)","volume":"50","author":"Amato","year":"2022","journal-title":"Toxicol. Pathol."},{"key":"ref_16","first-page":"233","article-title":"The global landscape of approved antibody therapies","volume":"5","author":"Lyu","year":"2022","journal-title":"Antib. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"256","DOI":"10.4161\/mabs.2.3.11641","article-title":"The immunogenicity of humanized and fully human antibodies: Residual immunogenicity resides in the CDR regions","volume":"2","author":"Harding","year":"2010","journal-title":"mAbs"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1298473","DOI":"10.1155\/2016\/1298473","article-title":"Immunogenicity of Biotherapeutics: Causes and Association with Posttranslational Modifications","volume":"2016","author":"Kuriakose","year":"2016","journal-title":"J. Immunol. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.21873\/anticanres.14062","article-title":"Systematic Review on Infusion Reactions to and Infusion Rate of Monoclonal Antibodies Used in Cancer Treatment","volume":"40","author":"Rombouts","year":"2020","journal-title":"Anticancer Res."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Boune, S., Hu, P., Epstein, A.L., and Khawli, L.A. (2020). Principles of N-Linked Glycosylation Variations of IgG-Based Therapeutics: Pharmacokinetic and Functional Considerations. Antibodies, 9.","DOI":"10.3390\/antib9020022"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1038\/nbt.1839","article-title":"Acceptable changes in quality attributes of glycosylated biopharmaceuticals","volume":"29","author":"Schiestl","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"704","DOI":"10.1080\/19420862.2017.1305530","article-title":"Drifts in ADCC-related quality attributes of Herceptin\u00ae: Impact on development of a trastuzumab biosimilar","volume":"9","author":"Kim","year":"2017","journal-title":"mAbs"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ejca.2019.07.015","article-title":"Three-year follow-up from a phase 3 study of SB3 (a trastuzumab biosimilar) versus reference trastuzumab in the neoadjuvant setting for human epidermal growth factor receptor 2-positive breast cancer","volume":"120","author":"Pivot","year":"2019","journal-title":"Eur. J. Cancer"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1820","DOI":"10.1158\/0008-5472.CAN-17-1958","article-title":"Fc-Mediated Anomalous Biodistribution of Therapeutic Antibodies in Immunodeficient Mouse Models","volume":"78","author":"Sharma","year":"2018","journal-title":"Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"629","DOI":"10.2967\/jnumed.121.262383","article-title":"Influence of Fc Modifications and IgG Subclass on Biodistribution of Humanized Antibodies Targeting L1CAM","volume":"63","author":"Sharma","year":"2022","journal-title":"J. Nucl. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"495","DOI":"10.1038\/256495a0","article-title":"Continuous cultures of fused cells secreting antibody of predefined specificity","volume":"256","author":"Kohler","year":"1975","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1038\/321522a0","article-title":"Replacing the complementarity-determining regions in a human antibody with those from a mouse","volume":"321","author":"Jones","year":"1986","journal-title":"Nature"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1016\/0022-2836(87)90412-8","article-title":"Canonical structures for the hypervariable regions of immunoglobulins","volume":"196","author":"Chothia","year":"1987","journal-title":"J. Mol. Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1119","DOI":"10.4049\/jimmunol.169.2.1119","article-title":"\u201cSuperhumanized\u201d antibodies: Reduction of immunogenic potential by complementarity-determining region grafting with human germline sequences: Application to an anti-CD28","volume":"169","author":"Tan","year":"2002","journal-title":"J. Immunol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1038\/nprot.2010.13","article-title":"Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine","volume":"5","author":"Vosjan","year":"2010","journal-title":"Nat. Protoc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.nucmedbio.2017.03.004","article-title":"Production of Zr-89 using sputtered yttrium coin targets 89Zr using sputtered yttrium coin targets","volume":"50","author":"Queern","year":"2017","journal-title":"Nucl. Med. Biol."},{"key":"ref_32","unstructured":"(2002). Basic Anatomical and Physiological Data for Use in Radiological Protection Reference Values. Ann. ICRP, 32, 5\u2013265."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/3\/295\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:39:42Z","timestamp":1760027982000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/18\/3\/295"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,21]]},"references-count":32,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["ph18030295"],"URL":"https:\/\/doi.org\/10.3390\/ph18030295","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,21]]}}}