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Multimodality Imaging of Coiled‐Coil Mediated Self‐Assembly in a “Drug‐Free” Therapeutic System
Author(s) -
Zhang Rui,
Yang Jiyuan,
Chu TeWei,
Hartley Jonathan M.,
Kopeček Jindřich
Publication year - 2015
Publication title -
advanced healthcare materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.288
H-Index - 90
eISSN - 2192-2659
pISSN - 2192-2640
DOI - 10.1002/adhm.201400679
Subject(s) - conjugate , confocal , confocal microscopy , cell , colocalization , drug delivery , biophysics , chemistry , materials science , cancer research , microbiology and biotechnology , medicine , biochemistry , nanotechnology , biology , mathematical analysis , geometry , mathematics
Two complementary coiled‐coil peptides CCE/CCK are used to develop a “drug free” therapeutic system, which can specifically kill cancer cells without a drug. CCE is attached to the Fab′ fragment of anti‐CD20 1F5 antibody (Fab′‐CCE), and CCK is conjugated in multiple grafts to poly[ N ‐(2‐hydroxypropyl)methacrylamide] (P‐(CCK) x ). Two conjugates are consecutively administered: First, Fab′‐CCE coats peptide CCE at CD20 antigen of lymphoma cell surface; second, CCE/CCK biorecognition between Fab′‐CCE and P‐(CCK) x leads to coiled‐coil formation, CD20 crosslinking, membrane reorganization, and ultimately cell apoptosis. To prove that two conjugates can assemble at cell surface, multiple fluorescence imaging studies are performed, including 2‐channel FMT, 3D confocal microscopy, and 4‐color FACS. Confocal microscopy shows colocalization of two fluorescently labeled conjugates on non‐Hodgkin's lymphoma (NHL) Raji cell surface, indicating “two‐step” targeting specificity. The fluorescent images also reveal that these two conjugates can disrupt normal membrane lipid distribution and form lipid raft clusters, leading to cancer cell apoptosis. This “two‐step” biorecognition capacity is further demonstrated in a NHL xenograft model, using fluorescent images at whole‐body, tissue and cell levels. It is also found that delaying injection of P‐(CCK) x can significantly enhance targeting efficacy. This high‐specificity therapeutics provide a safe option to treat NHL and other B cell malignancies.