Skip to main content
Top
Published in: European Journal of Nuclear Medicine and Molecular Imaging 1/2006

01-01-2006 | Original Article

99mTc-rituximab radiolabelled by photo-activation: a new non-Hodgkin’s lymphoma imaging agent

Authors: T. Gmeiner Stopar, I. Mlinaric-Rascan, J. Fettich, S. Hojker, S. J. Mather

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 1/2006

Login to get access

Abstract

Purpose

Rituximab was the first chimeric monoclonal antibody to be approved for treatment of indolent B-cell non-Hodgkin’s lymphoma (NHL). It is directed against the CD20 antigen, which is expressed by 95% of B-cell NHLs. The aim of this study was to explore the possibility of radiolabelling rituximab with 99mTc for use as an imaging agent in NHL for early detection, staging, remission assessment, monitoring for metastatic spread and tumour recurrence, and assessment of CD20 expression prior to (radio)immunotherapy.

Methods

Rituximab was purified from Mabthera solution (Roche), photo-activated at 302 nm by UV irradiation and radiolabelled with 99mTc. The effectiveness of the labelling method was evaluated by determination of the number of free thiol groups per photoreduced antibody, radiochemical purity and in vitro stability of 99mTc-rituximab.

Results

On average, 4.4 free thiol groups per photoreduced antibody were determined. Radiolabelling yields greater than 95% were routinely observed after storage of the photo-activated antibody at −80°C for 195 days. The direct binding assay showed preserved ability of 99mTc-rituximab to bind to CD20, with an average immunoreactive fraction of 93.3%. The internalisation rate was proven to be low, with only 5.3% of bound 99mTc-rituximab being internalised over 4 h at 37°C.

Conclusion

Our results demonstrate that 99mTc-rituximab of high radiochemical purity and with preserved binding affinity for the antigen can be prepared by photoreduction and that the method shows good reproducibility. 99mTc-rituximab will be further explored as an imaging agent applicable in NHL for the purposes mentioned above.
Literature
1.
go back to reference Golay J, Gramigna R, Facchinetti V, Capello D, Gaidano G, Introna M. Acquired immunodeficiency syndrome-associated lymphomas are efficiently lysed through complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity by rituximab. Br J Haematol 2002;119:923–9CrossRefPubMed Golay J, Gramigna R, Facchinetti V, Capello D, Gaidano G, Introna M. Acquired immunodeficiency syndrome-associated lymphomas are efficiently lysed through complement-dependent cytotoxicity and antibody-dependent cellular cytotoxicity by rituximab. Br J Haematol 2002;119:923–9CrossRefPubMed
2.
go back to reference Cragg MS, Walshe CA, Ivanov AO, Glennie MJ. The biology of CD20 and its potential as a target for mAb therapy. Curr Dir Autoimmun 2005;8:140–74PubMed Cragg MS, Walshe CA, Ivanov AO, Glennie MJ. The biology of CD20 and its potential as a target for mAb therapy. Curr Dir Autoimmun 2005;8:140–74PubMed
3.
go back to reference Coffey J, Hodgson DC, Gospodarowicz MK. Therapy of non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S28–36PubMed Coffey J, Hodgson DC, Gospodarowicz MK. Therapy of non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S28–36PubMed
4.
go back to reference Gopal AK, Press OW. Clinical applications of anti-CD20 antibodies. J Lab Clin Med 1999;134:445–50CrossRefPubMed Gopal AK, Press OW. Clinical applications of anti-CD20 antibodies. J Lab Clin Med 1999;134:445–50CrossRefPubMed
6.
go back to reference Withoff S, Bijman MNA, Stel AJ, Delahaye L, Calogero A, de Jonge MWA, et al. Characterization of BIS20x3, a bi-specific antibody activating and retargeting T-cells to CD20-positive B-cells. Br J Cancer 2001;84:1115–21CrossRefPubMed Withoff S, Bijman MNA, Stel AJ, Delahaye L, Calogero A, de Jonge MWA, et al. Characterization of BIS20x3, a bi-specific antibody activating and retargeting T-cells to CD20-positive B-cells. Br J Cancer 2001;84:1115–21CrossRefPubMed
7.
go back to reference Avivi I, Robinson S, Goldstone A. Clinical use of rituximab in haematological malignacies. Minireview. Br J Cancer 2003;89:1389–94CrossRefPubMed Avivi I, Robinson S, Goldstone A. Clinical use of rituximab in haematological malignacies. Minireview. Br J Cancer 2003;89:1389–94CrossRefPubMed
8.
10.
go back to reference Emmanouilides C. Current status of radioimmunotherapy for non-Hodgkin lymphoma. Haema 2003;6:314–27 Emmanouilides C. Current status of radioimmunotherapy for non-Hodgkin lymphoma. Haema 2003;6:314–27
11.
go back to reference Kaminski MS, Zasadny KR, Francis IR, Milik AW, Ross CW, Moon SD, et al. Radioimmunotherapy of B-cell lymphoma with (131I)anti-B1 (anti-CD20) antibody. N Engl J Med 1993;329:459–65CrossRefPubMed Kaminski MS, Zasadny KR, Francis IR, Milik AW, Ross CW, Moon SD, et al. Radioimmunotherapy of B-cell lymphoma with (131I)anti-B1 (anti-CD20) antibody. N Engl J Med 1993;329:459–65CrossRefPubMed
13.
go back to reference Swerdlow AJ. Epidemiology of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S2–12 Swerdlow AJ. Epidemiology of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S2–12
14.
go back to reference Hoskin PJ, O’Doherty MJ. Lymphoma: diagnosis and management introduction. Eur J Nucl Med Mol Imaging 2003; 30 Suppl 1:S1CrossRef Hoskin PJ, O’Doherty MJ. Lymphoma: diagnosis and management introduction. Eur J Nucl Med Mol Imaging 2003; 30 Suppl 1:S1CrossRef
15.
go back to reference Even-Sapir E, Israel O. Gallium-67 scintigraphy: a cornerstone in functional imaging of lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S65–81PubMed Even-Sapir E, Israel O. Gallium-67 scintigraphy: a cornerstone in functional imaging of lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S65–81PubMed
16.
go back to reference Hoskin PJ. PET in lymphoma: what are the oncologist’s needs? Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S37–41CrossRefPubMed Hoskin PJ. PET in lymphoma: what are the oncologist’s needs? Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S37–41CrossRefPubMed
17.
go back to reference Rankin SC. Assessment of response to therapy using conventional imaging. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S56–64PubMed Rankin SC. Assessment of response to therapy using conventional imaging. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S56–64PubMed
18.
go back to reference Sciepers C, Filmont JE, Czernin J. PET for staging of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S82–8PubMed Sciepers C, Filmont JE, Czernin J. PET for staging of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S82–8PubMed
19.
go back to reference Vinnicombe SJ, Reznek RH. Computerised tomography in the staging of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S42–55PubMed Vinnicombe SJ, Reznek RH. Computerised tomography in the staging of Hodgkin’s disease and non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S42–55PubMed
20.
go back to reference Reske SN. PET and restaging of malignant lymphoma including residual masses and relapse. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S82–96PubMed Reske SN. PET and restaging of malignant lymphoma including residual masses and relapse. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S82–96PubMed
21.
go back to reference Spaepen K, Stroobants S, Verhoef G, Mortelamns L. Positron emission tomography with [18F]FDG for therapy response monitoring in lymphoma patients. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S97–105PubMed Spaepen K, Stroobants S, Verhoef G, Mortelamns L. Positron emission tomography with [18F]FDG for therapy response monitoring in lymphoma patients. Eur J Nucl Med Mol Imaging 2003;30 Suppl 1:S97–105PubMed
22.
go back to reference Reilly RM. Immunoscintigraphy of tumours using 99mTc-labelled monoclonal antibodies: a review. Nucl Med Commun 1993;14:347–59PubMed Reilly RM. Immunoscintigraphy of tumours using 99mTc-labelled monoclonal antibodies: a review. Nucl Med Commun 1993;14:347–59PubMed
23.
go back to reference Mather SJ. Radiolabelled antibodies and peptides. In: Sampson CB, editor. Textbook of radiopharmacy theory and practice. 3rd ed. Amsterdam, The Netherlands: Gordon and Breach Science Publishers; 1990. p. 63–82 Mather SJ. Radiolabelled antibodies and peptides. In: Sampson CB, editor. Textbook of radiopharmacy theory and practice. 3rd ed. Amsterdam, The Netherlands: Gordon and Breach Science Publishers; 1990. p. 63–82
24.
go back to reference Urbain JLC, Vekemans MCM, Lemieux SK, Cosenza SC, Senadhi VK, Milestone BN, et al. Nuclear oncology and the imagine concept. Acta Radiol 1997;38 Suppl 41:21–8 Urbain JLC, Vekemans MCM, Lemieux SK, Cosenza SC, Senadhi VK, Milestone BN, et al. Nuclear oncology and the imagine concept. Acta Radiol 1997;38 Suppl 41:21–8
25.
go back to reference Stalteri MA, Mather SJ. Technetium-99m labelling of the anti-tumour antibody PR1A3 by photoactivation. Eur J Nucl Med 1996;23:178–87CrossRefPubMed Stalteri MA, Mather SJ. Technetium-99m labelling of the anti-tumour antibody PR1A3 by photoactivation. Eur J Nucl Med 1996;23:178–87CrossRefPubMed
26.
go back to reference Muddukrishna SN, Chen A, Qi P, Smolenski MA. Quantitation of reduced disulfide groups in monoclonal antibodies using 5-iodoacetamidofluorescein: a novel size exclusion-HPLC technique. Appl Radiat Isotopes 1995;46:1015–26CrossRef Muddukrishna SN, Chen A, Qi P, Smolenski MA. Quantitation of reduced disulfide groups in monoclonal antibodies using 5-iodoacetamidofluorescein: a novel size exclusion-HPLC technique. Appl Radiat Isotopes 1995;46:1015–26CrossRef
27.
go back to reference Lindmo T, Boven E, Cuttitta F, Fedorko J, Bunn PA Jr. Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods 1984;72:77–89CrossRefPubMed Lindmo T, Boven E, Cuttitta F, Fedorko J, Bunn PA Jr. Determination of the immunoreactive fraction of radiolabeled monoclonal antibodies by linear extrapolation to binding at infinite antigen excess. J Immunol Methods 1984;72:77–89CrossRefPubMed
28.
go back to reference Badger CC, Krohn KA, Bernstein ID. In vitro measurement of avidity of radioiodinated antibodies. Nucl Med Biol 1987;14:605–10 Badger CC, Krohn KA, Bernstein ID. In vitro measurement of avidity of radioiodinated antibodies. Nucl Med Biol 1987;14:605–10
29.
go back to reference Divgi CR. Status of radiolabeled monoclonal antibodies for diagnosis and therapy of cancer. Oncology 1996;10:939–58PubMed Divgi CR. Status of radiolabeled monoclonal antibodies for diagnosis and therapy of cancer. Oncology 1996;10:939–58PubMed
30.
go back to reference Juweid ME. Radiotherapy of B-cell non-Hodgkin’s lymphoma: from clinical trials to clinical practice. J Nucl Med 2002;43:1507–29PubMed Juweid ME. Radiotherapy of B-cell non-Hodgkin’s lymphoma: from clinical trials to clinical practice. J Nucl Med 2002;43:1507–29PubMed
31.
go back to reference Michel RB, Mattes MJ. Intracellular accumulation of the anti-CD20 antibody 1F5 in B-lymphoma cells. Clin Cancer Res 2002;8:2701–13PubMed Michel RB, Mattes MJ. Intracellular accumulation of the anti-CD20 antibody 1F5 in B-lymphoma cells. Clin Cancer Res 2002;8:2701–13PubMed
32.
go back to reference Press OW, Farr AG, Borroz KI, Anderson SK, Martin PJ. Endocytosis and degradation of monoclonal antibodies targeting human B-cell malignancies. Cancer Res 1989;49:4906–12PubMed Press OW, Farr AG, Borroz KI, Anderson SK, Martin PJ. Endocytosis and degradation of monoclonal antibodies targeting human B-cell malignancies. Cancer Res 1989;49:4906–12PubMed
33.
go back to reference Jilani I, O’Brien S, Manshuri T, Thomas DA, Thomazy VA, Imam M, et al. Transient down-modulation of CD20 by rituximab in patients with chronic lymphocytic leukemia. Blood 2003;102:3514–20CrossRefPubMed Jilani I, O’Brien S, Manshuri T, Thomas DA, Thomazy VA, Imam M, et al. Transient down-modulation of CD20 by rituximab in patients with chronic lymphocytic leukemia. Blood 2003;102:3514–20CrossRefPubMed
34.
go back to reference Schaffland AO, Buchegger F, Kosinski M, Antonescu C, Paschoud C, Grannavel C, et al. 131I-rituximab: relationship between immunoreactivity and specific activity. J Nucl Med 2004;45:1784–90PubMed Schaffland AO, Buchegger F, Kosinski M, Antonescu C, Paschoud C, Grannavel C, et al. 131I-rituximab: relationship between immunoreactivity and specific activity. J Nucl Med 2004;45:1784–90PubMed
35.
go back to reference Scheidhauer K, Wolf I, Baumgartl HJ, Von Schilling C, Schmidt B, Reidel G, et al. Biodistribution and kinetics of 131I-labelled anti-CD20 MAB IDEC-C2B8 (rituximab) in relapsed non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2002;29:1276–82CrossRefPubMed Scheidhauer K, Wolf I, Baumgartl HJ, Von Schilling C, Schmidt B, Reidel G, et al. Biodistribution and kinetics of 131I-labelled anti-CD20 MAB IDEC-C2B8 (rituximab) in relapsed non-Hodgkin’s lymphoma. Eur J Nucl Med Mol Imaging 2002;29:1276–82CrossRefPubMed
Metadata
Title
99mTc-rituximab radiolabelled by photo-activation: a new non-Hodgkin’s lymphoma imaging agent
Authors
T. Gmeiner Stopar
I. Mlinaric-Rascan
J. Fettich
S. Hojker
S. J. Mather
Publication date
01-01-2006
Publisher
Springer-Verlag
Published in
European Journal of Nuclear Medicine and Molecular Imaging / Issue 1/2006
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI
https://doi.org/10.1007/s00259-005-1838-4

Other articles of this Issue 1/2006

European Journal of Nuclear Medicine and Molecular Imaging 1/2006 Go to the issue