Poly l lysine hydrobromide pll

Poly-L-lysine hydrobromide (PLL) (MW range = 4000–15,000) and polyamidoamine (PAMAM) dendrimer with an ethylenediamine core (generation (G) 3) in methanol were purchased from Sigma-Aldrich Corp. (St. Louis, MO, USA). Boc-Ser(tBu)-OH and HOBt were purchased from Watanabe Chemical Industries (Hiroshima, Japan). Hexafluorophosphate benzotriazole tetramethyl uronium (HBTU) was purchased from Merck Millipore (Burlington, MA, USA). Dimethyl sulfoxide (DMSO; super dehydrated grade), N,N-dimethylformamide (DMF; super dehydrated grade), N,N-diisopropylethylamine (DIPEA), piperidine, trifluoroacetic acid (TFA), triisopropylsilane (TIS), Turk’s solution, and poly-L-lysine hydrobromide (MW range = 15,000–30,000) were purchased from Fujifilm Wako Pure Chemical Industries Ltd. (Osaka, Japan). PD-10 was purchased from GE Healthcare Japan (Tokyo, Japan). The ¹¹¹InCl₃ was supplied by Nihon Medi-Physics (Tokyo, Japan). ⁹⁰YCl₃ was purchased from Eckert Radiopharma (Berlin, Germany). DTPA anhydride was purchased from Chemical Dojin Co., Ltd. (Kumamoto, Japan). All other chemicals were of commercial reagent grade.

The pGluc vector (5764 bp) or pBMP2 vector (3486 bp) (100 μg) was complexed with 500 μg of Poly L-Lysine hydrobromide (PLL) (1000–5000 Da, Sigma, United Kingdom) by sequentially adding 10 μg of pDNA dropwise to 50 μg of PLL using a P200 pipette tip under vortexing. The working concentration was 40 and 200 μg/ml in nuclease-free water for pDNA and PLL, respectively. The mixture of pDNA-PLL NPs in a volume of 5 ml was then concentrated to 250 μl using Amicon^® Ultra Centrifugal Filter units. This volume of the mixture of pDNA-PLL NPs was encapsulated in the PLGA NPs by double emulsion as below. The hydrodynamic size and surface charge of pDNA-PLL NPs were assessed by Malvern Zetasizer Nano ZS. NP morphology was determined by Transmission Electron Microscopy (FEI Tecnai BioTwin-12 TEM) as previously (Osman et al., 2018 (link)).