Polyethyleneimine as a transmembrane carrier of fluorescently labeled proteins and antibodies

被引:32
作者
Didenko, VV [1 ]
Ngo, H
Baskin, DS
机构
[1] Baylor Coll Med, Houston, TX 77030 USA
[2] Michael E DeBakey Vet Affairs Med Ctr, Houston, TX 77030 USA
[3] Methodist Neurol Inst, Houston, TX 77030 USA
关键词
polyethyleneimine; PEI; protein delivery; avidin; Alexa avidin conjugates; lamin; glial cells; fibroblasts; antibody delivery; protein carrier; fluorescent detection;
D O I
10.1016/j.ab.2005.06.011
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Polyethyleneimine (PEI) has been used previously as a nonviral DNA transfer vector. In this article, we demonstrate its use as a vehicle for transmembrane delivery of proteins in cell culture conditions. Linking proteins to PEI required no other treatment beyond mixing them with PEI. The bond between PEI and protein combined at optimal ratios was maintained in electrophoresis, even in the presence of 2.5% sodium dodecyl sulfate (SIDS). The optimal time for delivery of proteins was determined to be 24 h. We have successfully delivered an Alexa 488-labeled avidin protein into human glioblastoma cells. A functional antibody against the nuclear protein lamin was delivered into human fibroblasts and reacted with lamin inside live cells. PEI-based delivery of antibodies and fluorescently labeled proteins can be used for fluorescent detection, tracking, and evaluation of cellular protein function in vivo. (C) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:168 / 173
页数:6
相关论文
共 12 条
[1]  
DEROSSI D, 1994, J BIOL CHEM, V269, P10444
[2]   Intercellular trafficking and protein delivery by a herpesvirus structural protein [J].
Elliott, G ;
OHare, P .
CELL, 1997, 88 (02) :223-233
[3]   TAT-MEDIATED DELIVERY OF HETEROLOGOUS PROTEINS INTO CELLS [J].
FAWELL, S ;
SEERY, J ;
DAIKH, Y ;
MOORE, C ;
CHEN, LL ;
PEPINSKY, B ;
BARSOUM, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (02) :664-668
[4]   IMMUNOCYTOCHEMICAL LOCALIZATION OF MAJOR POLYPEPTIDES OF NUCLEAR-PORE COMPLEX-LAMINA FRACTION - INTERPHASE AND MITOTIC DISTRIBUTION [J].
GERACE, L ;
BLUM, A ;
BLOBEL, G .
JOURNAL OF CELL BIOLOGY, 1978, 79 (02) :546-566
[5]   Poly(ethylenimine) and its role in gene delivery [J].
Godbey, WT ;
Wu, KK ;
Mikos, AG .
JOURNAL OF CONTROLLED RELEASE, 1999, 60 (2-3) :149-160
[6]   Tracking the intracellular path of poly(ethylenimine)/DNA complexes for gene delivery [J].
Godbey, WT ;
Wu, KK ;
Mikos, AG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (09) :5177-5181
[7]  
Harborth J, 2001, J CELL SCI, V114, P4557
[8]  
Haugland R.P., 2002, Handbook of Fluorescent Probes and Research Products, V9th
[9]  
Lemkine GF, 2001, CURR OPIN MOL THER, V3, P178
[10]   A peptide carrier for the delivery of biologically active proteins into mammalian cells [J].
Morris, MC ;
Depollier, J ;
Mery, J ;
Heitz, F ;
Divita, G .
NATURE BIOTECHNOLOGY, 2001, 19 (12) :1173-1176