2001
DOI: 10.1021/bc0001488
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Different Strategies for Formation of PEGylated EGF-Conjugated PEI/DNA Complexes for Targeted Gene Delivery

Abstract: With the aim of generating gene delivery systems for tumor targeting, we have synthesized a conjugate consisting of polyethylenimine (PEI) covalently modified with epidermal growth factor (EGF) peptides. Transfection efficiency of the conjugate was evaluated and compared to native PEI in three tumor cell lines: KB epidermoid carcinoma cells, CMT-93 rectum carcinoma cells, and Renca-EGFR renal carcinoma cells. Depending on the tumor cell line, incorporation of EGF resulted in an up to 300-fold increased transfe… Show more

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Cited by 235 publications
(175 citation statements)
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References 52 publications
(68 reference statements)
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“…(Peng et al, 2001;Wolschek et al, 2002;Ogris et al, 2003;Pereboeva et al, 2007). In our own work, we have most often used murine EGF, which, in contrast to human EGF, does not carry any lysines in its primary amino acid sequence (Blessing et al, 2001;Wolschek et al, 2002;Ogris et al, 2003). Hence, convenient coupling via the N-terminal primary amine to heterobifunctional cross-linkers, such as N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), can occur without interfering with the binding domain of EGF.…”
Section: Discussionmentioning
confidence: 99%
“…(Peng et al, 2001;Wolschek et al, 2002;Ogris et al, 2003;Pereboeva et al, 2007). In our own work, we have most often used murine EGF, which, in contrast to human EGF, does not carry any lysines in its primary amino acid sequence (Blessing et al, 2001;Wolschek et al, 2002;Ogris et al, 2003). Hence, convenient coupling via the N-terminal primary amine to heterobifunctional cross-linkers, such as N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP), can occur without interfering with the binding domain of EGF.…”
Section: Discussionmentioning
confidence: 99%
“…27 However, to date, most targeting sterically stabilized gene delivery vehicles have been studied in vitro. [3][4][5][6][7][8][9] As the ultimate goal of any potential gene therapy is delivery in vivo, a comparison study of TsPLP both in vitro and in vivo was undertaken to elucidate the influence of the PEGylation on gene delivery efficiency. PLP showed the lowest in vitro transfection activity (Figure 2), and this is likely due to the PEG molecule preventing association of the lipoplex with the target cell 2,12 and/or due to PEG inhibition of fusion between lipid and endosomal membrane, which is necessary for the intracellular release of exogenously delivered DNA.…”
Section: Discussionmentioning
confidence: 99%
“…In this study, postcoated TsPLP was prepared essentially according to a strategy described by Blessing et al 3 Briefly, DNA was mixed with liposomes composed of DOTAP and DOPE (1:1 molar ratio) to form a lipid/DNA complex (lipoplex). Subsequently, heterobifunctional PEG (NHS-PEG-MAL) was added to form a Mal-PEGlipoplex, in which a coupling reaction occurs between the NHS group of bifunctional PEG and the amine residue of DOPE (Figure 1).…”
Section: In Vitro Transfection Activitiesmentioning
confidence: 99%
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“…Intravenous injection of a PEGylated polyplex would result in passive accumulation of the polyplex in a tumor. However, to advance the beneficial effects of gene therapies, researchers have begun to target polycations in a noninvasive but directed manner 7,8 by complexing polycations with antibodies, vitamins, glycolipids, and receptor ligands, like transferrin (Tf).…”
Section: Introductionmentioning
confidence: 99%