Document Detail

A general strategy for generating intact, full-length IgG antibodies that penetrate into the cytosol of living cells.
MedLine Citation:
PMID:  25484049     Owner:  NLM     Status:  Publisher    
Abstract Full-length IgG antibodies cannot cross cell membranes of living cells; this limits their use for direct targeting of cytosolic proteins. Here, we describe a general strategy for the generation of intact, full-length IgG antibodies, herein called cytotransmabs, which internalize into living cells and localize in the cytosol. We first generated a humanized light chain variable domain (VL) that could penetrate into the cytosol of living cells and was engineered for association with various subtypes of human heavy chain variable domains (VHs). When light chains with humanized VL were co-expressed with three heavy chains (HCs), including two HCs of the clinically approved adalimumab (Humira®) and bevacizumab (Avastin®), all three purified IgG antibodies were internalized into the cytoplasm of living cells. Cytotransmabs primarily internalized into living cells by the clathrin-mediated endocytic pathway through interactions with heparin sulfate proteoglycan that was expressed on the cell surface. The cytotransmabs escaped into the cytosol from early endosomes without being further transported into other cellular compartments, like the lysosomes, endoplasmic reticulum, Golgi apparatus, and nucleus. Furthermore, we generated a cytotransmab that co-localized with the targeted cytosolic protein when it was incubated with living cells, demonstrating that the cytotransmab can directly target cytosolic proteins. Internalized cytotransmabs did not show any noticeable cytotoxicity and remained in the cytosol for more than 6 h before being degraded by proteosomes. These results suggest that cytotransmabs, which efficiently enter living cells and reach the cytosolic space, will find widespread uses as research, diagnostic, and therapeutic agents.
Dong-Ki Choi; Jeomil Bae; Seung-Min Shin; Ju-Yeon Shin; Sunghoon Kim; Yong-Sung Kim
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-11-1
Journal Detail:
Title:  mAbs     Volume:  -     ISSN:  1942-0870     ISO Abbreviation:  MAbs     Publication Date:  2014 Nov 
Date Detail:
Created Date:  2014-12-8     Completed Date:  -     Revised Date:  2014-12-9    
Medline Journal Info:
Nlm Unique ID:  101479829     Medline TA:  MAbs     Country:  -    
Other Details:
Languages:  ENG     Pagination:  0     Citation Subset:  -    
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