Because antibodies cannot enter cells, past experiments using antibodies focused primarily on extracellular molecules.If target molecules in living cells can be exposed to antibodies, it will be possible to pursue new dimensions of research related to cell function analysis, exploration of target molecules for disease diagnosis and treatment, and so on. GenomONE-CAb Antibody Delivery Reagent is a next-generation tool for antibody introduction into cells which can meet such needs.With this kit, antibody can be incorporated into the HVJ Envelope (HVJ-E), a transfection tool making use of the membrane fusing ability of inactivated Sendai virus (HVJ: Hemagglutinating Virus of Japan). If cells are treated with HVJ-E including antibody, it will be possible to achieve efficient introduction of IgG antibodies into the cytoplasm. This kit provides a totally new methodology for experiments, overcoming the difficulties involved in experiments using conventional lipid-based reagents by which antibodies are introduced into cells by means of endocytosis.
Experimental Protocol
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Principle
What is HVJ Envelope(HVJ-E)?
With this system, the IgG antibody incorporation enhancer, which was developed by our company, improves the efficiency of inclusion of IgG molecules into HVJ-E markedly compared to the existing HVJ-E vectors (GenomONE, GenomONE-Neo ) marketed in 2002. Thanks to this feature, this system allows efficient introduction of IgG molecules into the cytoplasm.
<Reference (Published research article) >
Efficient delivery of antibody into living cells using a novel HVJ envelope vector system.
Y. Kondo et al., J. Immunol. Methods, 332, 10-17 (2008)
Features
Analysis of intracellular function
- An antibody is introduced into living cells to examine the distribution of the target molecule within them
- An antibody is introduced into living cells to suppress and clarify the function of target molecules
- Live cell imaging is performed
Screening of antibodies reacting to intracellular antigens
- Antibodies binding to intact antigens in living cells and showing neutralization activity are screened for
Application to testing, diagnosis, and treatment
- New agents for testing and diagnosis are developed using antibodies capable of detecting target molecules in living cells
- Next-generation antibody-based drugs are created, which exert therapeutic effects through acting on intracellular target molecules
Advantages of intracellular antibody introduction (differences from existing knockout method)
- Unlike post-transcriptional gene silencing (RNAi method, etc.), this method is expected to achieve specific inhibition by recognizing protein-protein interactions or post-translational modifications (addition of sugar chains, etc.)
- Nonspecific reactions (off-target effects of RNAi method, etc.) are unlikely to occur
- Unlike gene transfer and expression methods, introduction of antibody in amounts sufficient to exert efficacy can be achieved rapidly and simply, and this method is applicable to a wider range of types of experiments
