Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 389, Issue 1-2, Pages 45-51Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2012.12.009
Keywords
Antibody; CpG oligodeoxynucleotides; Immunoconjugates; Immunotherapy; Rituximab; Trastuzumab
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Funding
- Cancer Therapeutics Laboratories, Inc. (Los Angeles, CA)
- Department of Defense [W81XWH-11-1-0466]
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A number of monoclonal antibodies against tumor-associated antigens have been developed for the treatment of cancer. The anti-tumor effects of such antibodies can be enhanced by conjugation to immune stimulatory ligands, such as the toll-like receptor 9 agonist CpG oligodeoxynucleotides (CpG). The present study describes methods for the conjugation of CpG to two clinically approved monoclonal antibodies (rituximab and trastuzumab) via a Sulfo-EMCS maleimide linker. This conjugation method yielded stable joining of CpG and antibody (molar range 2.2-4.3:1). Immunofluorescence studies showed intact antigen-specific antibody binding of the immunoconjugates, that were comparable to unmodified antibody. Furthermore, antibody-CpG conjugates demonstrated improved (rituximab) or equivalent (trastuzumab) immune stimulatory activity compared to free CpG in vitro. These studies demonstrate the feasibility of antibody-CpG immunoconjugates and provide the foundation for future in vivo immunotherapy evaluation. (C) 2012 Elsevier B.V. All rights reserved.
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