title: Pullulan-protamine as efficient haemocompatible gene delivery vector: synthesis and in vitro characterization.
abstract: Biodegradable non-viral vectors with good transfection efficiency is essential for successful gene delivery. The purpose of this study was to design a non-viral vector by conjugating protamine to pullulan and elucidate the potential use of pullulan protamine conjugate (PPA) as an effective, non toxic and haemocompatible gene delivery system. The particle size and surface charge were measured using Nanosizer. Derivatization was confirmed by NMR, FTIR and DSC analyses. Acid base titration revealed the buffering behaviour of the conjugate. The protection of DNA from nuclease enzyme and interaction of plasma components on the stability of nanoplexes were also analysed. The uptake studies confirmed the plasmid delivery into the nucleus and the inhibitor studies determined the uptake mechanism. Transfection experiments revealed the capability of PPA to cellular uptake in C6 cells and facilitate high gene expression. Thus, PPA proves to be a promising non-viral vector.
authors: Biosurface Technology Division, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala, India.
authors: S S
authors: SS
authors: Priya
authors: Biosurface Technology Division, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala, India. Electronic address: rekhamr@sctimst.ac.in.
authors: M R
authors: MR
authors: Rekha
authors: Biosurface Technology Division, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala, India. Electronic address: sharmacp@sctimst.ac.in.
authors: Chandra P
authors: CP
authors: Sharma
mesh_terms: D001672:Biocompatible Materials; D002152:Calorimetry, Differential Scanning; D004247:DNA / Q000008:administration & dosage; D005822:Genetic Vectors; D005936:Glucans / Q000008:administration & dosage / Q000737:chemistry*; D006461:Hemolysis; D066298:In Vitro Techniques; D009682:Magnetic Resonance Spectroscopy; D063347:Native Polyacrylamide Gel Electrophoresis; D010974:Platelet Aggregation; D011479:Protamines / Q000008:administration & dosage / Q000737:chemistry*; D017550:Spectroscopy, Fourier Transform Infrared; D014162:Transfection
chemical_list: D001672:Biocompatible Materials; D005936:Glucans; D011479:Protamines; C009109:pullulan; D004247:DNA
keywords: Cationized pullulan; Gene delivery; Haemocompatible; Non- viral vector; Protamine