title: Cationized bovine serum albumin as gene carrier: Influence of specific secondary structure on DNA complexibility and gene transfection.
abstract: In this research, BSA, one of the natural rigid globular proteins with ca. 51% of α-helix secondary structure, was utilized to prepare cationized BSA (cBSA) as gene carrier. Tetraethylenepentamine (TEPA) or polyethylenimine (PEI1800) was grafted to BSA with different grafting levels. Based on the circular dichoism (CD) spectra, all cBSA remained α-helical structure to some degree. This was exciting to endow cBSA with quite different DNA complexibility and cellular biology behavior from the random coiled and flexible polycations such as PEI and poly-l-lysine (PLL). Strangely, the DNA condensability decreased with the increment of TEPA or PEI1800 grafting level. Also, the cBSA could condense DNA effectively to form irregular nanoparticles around 50-200nm above N/P ratio of 10. On account of the excellent hydration of BSA, the cBSA/DNA complexes revealed good colloidal stability under physiological salt condition. Cell culture experiments indicated this BSA-based gene carrier possessed good cellular compatibility. Surprisingly, cBSA/DNA complexes could be uptaken excellently by up to 90% cells. This might be owing to the agitation effect of α-helical structure and the positive potential of these complexes. BSA-PEI1800/DNA complexes with quick endosome escape even had transfection efficiency as high as PEI25k/DNA complexes. Overall, this paper provided us the potential of cBSA as gene carrier and might have some instructions in the design of protein-based gene delivery system.
authors.affiliation: MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, PR China.
authors.forename: Jianwei
authors.initials: J
authors.lastname: Du
authors.affiliation: MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, PR China.
authors.forename: Bangbang
authors.initials: B
authors.lastname: Li
authors.affiliation: MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, PR China. Electronic address: yx_wang@zju.edu.cn.
authors.forename: Youxiang
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D002417:Cattle; D003102:Colloids; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D005029:Ethylenediamines / Q000737:chemistry*; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D011094:Polyethyleneimine / Q000737:chemistry*; D017433:Protein Structure, Secondary; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D055672:Static Electricity
chemical_list: D003102:Colloids; D005029:Ethylenediamines; D012710:Serum Albumin, Bovine; D011094:Polyethyleneimine; D004247:DNA; C034269:tetraethylenepentamine
keywords: Cationized BSA; Gene delivery; Hydration; Secondary structure