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Novel Protein Transfection of Primary Rat Cortical Neurons Using an Antibody That Penetrates Living Cells
Author(s) -
Richard H. Weisbart,
Roger A. Baldwin,
Brian Huh,
Debra Zack,
Robert N. Nishimura
Publication year - 2000
Publication title -
the journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.737
H-Index - 372
eISSN - 1550-6606
pISSN - 0022-1767
DOI - 10.4049/jimmunol.164.11.6020
Subject(s) - conjugate , catalase , cytoplasm , nucleus , hydrogen peroxide , transfection , intracellular , chemistry , microbiology and biotechnology , monoclonal antibody , fluorescence microscope , antibody , biophysics , biochemistry , fluorescence , biology , enzyme , immunology , mathematical analysis , physics , mathematics , quantum mechanics , gene
An Ab-based system to deliver functional proteins into neurons was developed using the murine mAb, mAb 3E10. This was achieved by covalently conjugating catalase to the Ab so that the conjugate retained high activity for the degradation of hydrogen peroxide. Three-dimensional fluorescence microscopy was used to demonstrate penetration of the Ab into the nucleus of living primary cortical neurons. The Ab conjugate localized in both the cytoplasm and nucleus. Retention of catalase activity after penetration and distribution of conjugate was demonstrated by reduction in cell death following exposure of treated neurons to hydrogen peroxide. These studies illustrate the potential of this method for the intracellular delivery of therapeutic proteins.

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