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64Cu-1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid-HB22.7

, PhD
National Center for Biotechnology Information, NLM, NIH, Bethesda, MD
Corresponding author.

Created: ; Last Update: May 15, 2009.

Chemical name:64Cu-1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid-HB22.7
Abbreviated name:64Cu-DOTA-HB22.7
Agent category:Antibody
Target category:Antigen
Method of detection:Positron emission tomography (PET)
Source of signal:64Cu
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Click on protein and gene for more information about the CD22.



CD22 is a transmembrane glycoprotein found on B lymphocytes. It functions as an adhesion molecule binding to sialic acid–bearing ligands with an immunoglobulin (Ig) domain and also modulates signal transduction via the B-cell receptor (1). CD22 is expressed on the cell surface of mature B cells and non-Hodgkin’s lymphomas (2). HB22.7, an anti-CD22 monoclonal antibody (mAb), blocks the interaction of the Ig domain of CD22 with its ligands to induce proliferation in primary B cells and apoptosis in neoplastic B cells (3, 4). 1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) has been successfully coupled to HB22.7 and labeled with 64Cu to produce a potential molecular imaging agent to target non-Hodgkin’s lymphomas (5). 64Cu is a positron emitter with a physical half-life of 12.7 h and is suitable for positron emission tomography (PET) imaging.



Martin et al. (5) reported the synthesis of 64Cu-DOTA-HB22.7. The HB22.7 mAb (67 nM) was incubated with 670 nM DOTA-NHS-ester in 0.1 M tetramethyl ammonium phosphate (pH 8.0) for 2.5 h at 37°C. DOTA-HB22.7 was isolated with column filtration. There were 6.3 DOTA molecules per antibody molecule. A mixture of DOTA-HB22.7 and 64Cu in 0.25 M ammonium acetate was incubated at 40°C for 40–60 min. 64Cu-DOTA-HB22.7 was purified with column filtration with a radiochemical yield of 83%. The radiochemical purity was >99% with a specific activity of 56 MBq/nmol (1.5 mCi/nmol) at the time of injection.

In Vitro Studies: Testing in Cells and Tissues


Martin et al. (5) performed flow cytometric analysis with DOTA-HB22.7 and HB22.7 using the CD22-expressing human Raji lymphoma cell line with a fluorescent-labeled anti-IgG as the secondary antibody. Binding of DOTA-HB22.7 and HB22.7 to Raji cells exhibited the same mean fluorescent intensity of 110 units, indicating that the immunoreactivity of DOTA-HB22.7 was similar to the parent mAb.

Animal Studies



Martin et al. (5) performed PET imaging studies in nude mice (n = 3/group) bearing Raji tumor xenografts. Each mouse received 7.4 MBq (0.2 mCi) 64Cu-DOTA-HB22.7 by intravenous injection. The radioactivity levels were obtained at 3 h, 24 h, and 48 h after injection, with tumor uptake values in percent injected dose per cc (% ID/cc) of 1.8, 5.5, and 6.8, respectively; tumor retention was high. Intraperitoneal and subcutaneous injections showed similar tumor uptakes at 24 and 48 h. Biodistribution studies at 48 h showed that the tumor (~18% ID/g) and blood (17–24% ID/g) exhibited the highest radioactivity levels for the three routes of injections. The distribution patterns to other nontargeted organs were similar with the exception of the spleen (14% ID/g), which showed the highest uptake with the intraperitoneal route compared with the other two routes (8% ID/g). The liver, kidneys, heart, lungs, and urinary bladder had similar radioactivity levels of ~5% ID/g. No blocking experiment was performed.

Other Non-Primate Mammals


No publication is currently available.

Non-Human Primates


No publication is currently available.

Human Studies


No publication is currently available.

NIH Support

R24 CA86307, R24 CA110804, R33 CA89706


Tedder T.F., Tuscano J., Sato S., Kehrl J.H. CD22, a B lymphocyte-specific adhesion molecule that regulates antigen receptor signaling. Annu Rev Immunol. 1997;15:481–504. [PubMed: 9143697]
Haas K.M., Sen S., Sanford I.G., Miller A.S., Poe J.C., Tedder T.F. CD22 ligand binding regulates normal and malignant B lymphocyte survival in vivo. J Immunol. 2006;177(5):3063–73. [PubMed: 16920943]
Tedder T.F., Poe J.C., Haas K.M. CD22: A Multifunctional Receptor That Regulates B Lymphocyte Survival and Signal Transduction. Adv Immunol. 2005;88:1–50. [PubMed: 16227086]
Tuscano J.M., Riva A., Toscano S.N., Tedder T.F., Kehrl J.H. CD22 cross-linking generates B-cell antigen receptor-independent signals that activate the JNK/SAPK signaling cascade. Blood. 1999;94(4):1382–92. [PubMed: 10438726]
Martin, S.M., R.T. O'Donnell, D.L. Kukis, C.K. Abbey, H. McKnight, J.L. Sutcliffe, and J.M. Tuscano, Imaging and Pharmacokinetics of (64)Cu-DOTA-HB22.7 Administered by Intravenous, Intraperitoneal, or Subcutaneous Injection to Mice Bearing Non-Hodgkin's Lymphoma Xenografts. Mol Imaging Biol, 2008. [PubMed: 18949521]


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