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TAT-LHRH conjugated low molecular weight chitosan as a gene carrier specific for hepatocellular carcinoma cells.

Liu L, Dong X, Zhu D, Song L, Zhang H, Leng XG - Int J Nanomedicine (2014)

Bottom Line: To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC).In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System.The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People's Republic of China.

ABSTRACT
To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.

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Characterization of synthesized TAT-LHRH peptide by (A) mass spectrometry and (B) high performance liquid chromatography.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; Intens, intensity; AU, arbitrary units; M/Z, mass-to-charge ratio.
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f1-ijn-9-2879: Characterization of synthesized TAT-LHRH peptide by (A) mass spectrometry and (B) high performance liquid chromatography.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; Intens, intensity; AU, arbitrary units; M/Z, mass-to-charge ratio.

Mentions: The synthesized TAT-Cys-LHRH peptide was analyzed with HPLC mass spectrometry to guarantee that a peak visible in the HPLC profile corresponds to the target mass. As shown in Figure 1, mass spectrometry and HPLC analysis demonstrated that the purity of the synthesized peptide was up to 99%, and the molecular mass was 2,657 daltons, which was identical to the expected molecular mass of the target peptide.


TAT-LHRH conjugated low molecular weight chitosan as a gene carrier specific for hepatocellular carcinoma cells.

Liu L, Dong X, Zhu D, Song L, Zhang H, Leng XG - Int J Nanomedicine (2014)

Characterization of synthesized TAT-LHRH peptide by (A) mass spectrometry and (B) high performance liquid chromatography.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; Intens, intensity; AU, arbitrary units; M/Z, mass-to-charge ratio.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061174&req=5

f1-ijn-9-2879: Characterization of synthesized TAT-LHRH peptide by (A) mass spectrometry and (B) high performance liquid chromatography.Abbreviations: TAT-LHRH, transactivator of transcription – luteinizing hormone-releasing hormone; Intens, intensity; AU, arbitrary units; M/Z, mass-to-charge ratio.
Mentions: The synthesized TAT-Cys-LHRH peptide was analyzed with HPLC mass spectrometry to guarantee that a peak visible in the HPLC profile corresponds to the target mass. As shown in Figure 1, mass spectrometry and HPLC analysis demonstrated that the purity of the synthesized peptide was up to 99%, and the molecular mass was 2,657 daltons, which was identical to the expected molecular mass of the target peptide.

Bottom Line: To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC).In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System.The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin, People's Republic of China.

ABSTRACT
To develop a chitosan-based nonviral gene carrier capable of delivering genes specifically into hepatoma cells, a bifunctional peptide composed of the TAT (transactivator of transcription) peptide and luteinizing hormone-releasing hormone (LHRH) was conjugated with low molecular weight chitosan, resulting in a TAT-LHRH-chitosan conjugate (TLC). TLC/DNA nanoparticles (TLCDNPs) were characterized by agarose gel retardation, atomic force microscopy, and dynamic light scattering analysis. In vitro targeting specificity and transfection efficiency were analyzed with a GE IN Cell Analyzer 2000 High-Content Cellular Analysis System. The results demonstrated that TLC had stronger DNA condensing power than unmodified chitosan, and that TLCDNPs were of roughly round shape with average diameter of 70-85 nm and zeta potential of +30 mV and were relatively stable in solution. The in vitro study demonstrated TLC was highly selective for hepatoma cells and essentially nontoxic.

Show MeSH
Related in: MedlinePlus