Human serum albumin (HSA), the most abundant protein in blood plasma, has been used as a drug carrier for the last few decades. Residualizingly radiolabeled serum albumin has been reported to be avidly taken up by tumors of sarcoma-bearing mice and to most likely undergo lysosomal degradation. In this study, we prepared 64Cu-1,4,7,10-tetraazacyclododecane-N,N′,N″,N′″-tetraacetic acid (DOTA) and Cy5.5-conjugated HSA (dual probe), and evaluated its tumor uptake and catabolism. Two dual probes were prepared using different DOTA conjugation sites of HSA (one via Lys residues and the other via the Cys residue). 64Cu-DOTA-Lys-HSA-Cy5.5 (dual probe-Lys) exhibited higher uptake by RR1022 sarcoma cells in vitro than 64Cu-DOTA-Cys-HSA-Cy5.5 (dual probe-Cys). In RR1022 tumor-bearing mice, the two dual probes showed a similar level of tumor uptake, but uptake of dual probe-Lys was reduced in the liver and spleen compared to dual probe-Cys, probably because of the presence of a higher number of DOTA molecules in the former. At 24 and 48 h after injection, dual probe-Lys was intact or partially degraded in blood, liver, kidney, and tumor samples, but 64Cu-DOTA-Lys was observed in the urine using radioactivity detection. Similarly, Cy5.5-Lys was observed in the urine using fluorescence detection. These results indicate that dual probe-Lys may be useful for predicting the catabolic fate of drug–HSA conjugates.
Human serum albumin PET/optical imaging 64Cu-DOTA-Lys Cy5.5-Lys Tumor Catabolism
This is a preview of subscription content, log in to check access.
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (grant code: 2011-0030164).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing financial interests.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The animal study was reviewed and approved by the Institutional Animal Care and Use Committee of SBRI. SBRI is an Association for Assessment and Accreditation of Laboratory Animal Care International accredited facility and abides by the Institute of Laboratory Animal Resources Guide.
This article does not contain any studies with human participants performed by any of the authors.
Andersson C, Iresjö BM, Lundholm K (1991) Identification of tissue sites for increased albumin degradation in sarcoma-bearing mice. J Surg Res 50:156–162CrossRefPubMedGoogle Scholar
Arano Y, Mukai T, Uezono T, Wakisaka K, Motonari H, Akizawa H, Taoka Y, Yokoyama A (1994) A biological method to evaluate bifunctional chelating agents to label antibodies with metallic radionuclides. J Nucl Med 35:890–898PubMedGoogle Scholar
Cai W, Wu Y, Chen K, Cao Q, Tice DA, Chen X (2006a) In vitro and in vivo characterization of 64Cu-labeled Abegrin™, a humanized monoclonal antibody against integrin αvβ3. Cancer Res 66:9673–9681CrossRefPubMedGoogle Scholar
Cai W, Chen K, Mohamedali KA, Cao Q, Gambhir SS, Rosenblum MG, Chen X (2006b) PET of vascular endothelial growth factor receptor expression. J Nucl Med 47:2048–2056PubMedGoogle Scholar
Chen K, Xie J, Chen X (2009) RGD-human serum albumin conjugates as efficient tumor targeting probes. Mol Imaging 8:65–73PubMedGoogle Scholar
de Rosales RTM, Tavaré R, Paul RL, Jauregui-Osoro M, Protti A, Glaria A, Varma G, Szanda I, Blower PJ (2011) Synthesis of 64CuII–bis(dithiocarbamatebisphosphonate) and its conjugation with superparamagnetic iron oxide nanoparticles: in vivo evaluation as dual-modality PET-MRI agent. Angew Chem Int Ed 50:5509–5513CrossRefGoogle Scholar
Elsadek B, Kratz F (2012) Impact of albumin on drug delivery—new application on the horizon. J Control Release 157:4–28CrossRefPubMedGoogle Scholar
Green MR, Manikhas GM, Orlov S, Afanasyev B, Makhson AM, Bhar P, Hawkins MJ (2006) Abraxane®, a novel Cremophor®-free, albumin-bound particle form of paclitaxel for the treatment of advanced non-small-cell lung cancer. Ann Oncol 17:1263–1268CrossRefPubMedGoogle Scholar
Grünberg J, Jeger S, Sarko D, Dennler P, Zimmermann K, Mier W, Schibli R (2013) DOTA functionalized polylysine: a high number of DOTA chelates positively influences the biodistribution of enzymatic conjugated anti-tumor antibody chCE7agl. PLoS ONE 8:e60350CrossRefPubMedPubMedCentralGoogle Scholar
Hackett MJ, Joolakanti S, Hartranft ME, Guley PC, Cho MJ (2012) A dicarboxylic fatty acid derivative of paclitaxel for albumin-assisted drug delivery. J Pharmaceut Sci 101:3292–3304CrossRefGoogle Scholar
Kang CM, An GI, Choe YS (2015) Hybrid lymph node imaging using 64Cu-labeled mannose-conjugated human serum albumin with and without indocyanine green. Nucl Med Commun 36:1026–1034PubMedGoogle Scholar
Kratz F (2011) INNO-206 (DOXO-EMCH), an albumin-binding prodrug of doxorubicin under development for phase II studies. Curr Bioact Compd 7:33–38CrossRefGoogle Scholar
Maxwell JL, Terracio Borg TK, Baynes JW, Thorpe SR (1990) A fluorescent residualizing label for studies on protein uptake and catabolism in vivo and in vitro. Biochem J 267:155–162CrossRefPubMedPubMedCentralGoogle Scholar
Meares CF, McCall MJ, Reardan DT, Goodwin DA, Diamanti CI, McTigue M (1984) Conjugation of antibodies with bifunctional chelating agents: isothiocyanate and bromoacetamide reagents, methods of analysis, and subsequent addition of metal ions. Anal Biochem 142:68–78CrossRefPubMedGoogle Scholar
Ono M, Arano Y, Uehara T, Fujioka Y, Ogawa K, Namba S, Mukai T, Nakayama M, Saji H (1999) Intracellular metabolic fate of radioactivity after injection of technetium-99 m-labeled hydrazino nicotinamide derivatized proteins. Bioconjugate Chem 10:386–394CrossRefGoogle Scholar
Peters T (1996) All about albumin: biochemistry, genetics, and medical applications. Academic Press, San DiegoGoogle Scholar
Rogers BE, Franano FN, Duncan JR, Edwards WB, Anderson CJ, Connett JM, Welch MJ (1995) Identification of metabolites of 111In-diethylenetriaminepentaacetic acid-monoclonal antibodies and antibody fragments in vivo. Cancer Res 55:5714s–5720sPubMedGoogle Scholar
Stehle G, Sinn H, Wunder A, Schrenk HH, Stewart JC, Hartung G, Maier-Borst W, Heene DL (1997) Plasma protein (albumin) catabolism by the tumor itself-implications for tumor metabolism and the genesis of cachexia. Crit Rev Oncol Hematol 26:77–100CrossRefPubMedGoogle Scholar
Thorpe SR, Baynes JW, Chroneos ZC (1993) The design and application of residualizing labels for studies of protein catabolism. FASEB J 7:399–405PubMedGoogle Scholar
Willner D, Trail PA, Hofstead SJ, King HD, Lasch SJ, Braslawsky GR, Greenfield RS, Kaneko T, Firestone RA (1993) (6-Maleimidocaproyl) hydrazone of doxorubicin-a new derivative for the preparation of immunoconjugates of doxorubicin. Bioconjugete Chem 4:521–527CrossRefGoogle Scholar
Wunder A, Stehle G, Sinn H, Schrenk HH, Hoff-Biederbeck D, Bader F, Friedrich EA, Peschke P, Maier-Borst W, Heene DL (1997) Enhanced albumin uptake by rat tumors. Int J Oncol 11:497–507PubMedGoogle Scholar
Yang W, Mou T, Peng C, Wu Z, Zhang X, Li F, Ma Y (2009) Fluorine-18 labeled galactosyl-neoglycoalbumin for imaging the hepatic asialoglycoprotein receptor. Bioorg Med Chem 17:7510–7516CrossRefPubMedGoogle Scholar
Yhee JY, Lee J, Chang H, Lee J, Kwon IC, Kim K (2015) Molecular imaging and targeted drug delivery using albumin-based nanoparticles. Curr Pharm Des 21:1889–1898CrossRefPubMedGoogle Scholar