Molar-Mass Characterization of Cationic Polymers for Gene Delivery by Aqueous Size-Exclusion Chromatography

Jiang, Xulin; Horst, Astrid van der; Steenbergen, Mies J. van; Akeroyd, Niels; Nostrum, Cornelus F. van; Schoenmakers, Peter J.; Hennink, Wim E.

doi:10.1007/s11095-006-9574-4

Cited by 63 publications

(57 citation statements)

References 41 publications

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“…A sodium acetate (NaAc) solution was prepared by dissolving a calculated amount of sodium acetate in reverse osmosis water (0.3 M), and the pH was adjusted to 4.4 with acetic acid. [39] A mixture of 70% 0.3 M NaAc solution, pH ¼ 4.4 and 30% acetonitrile was used as the eluent at a flow rate of 0.6 mL Á min À1 . The eluent was filtrated through a 0.22 mm HPLC filter and degassed prior to use by ultrasound.…”

Section: Synthesis Of Pei-ss-hp (7) Via Click Reactionmentioning

confidence: 99%

Disulfide‐Containing Hyperbranched Polyethylenimine Derivatives via Click Chemistry for Nonviral Gene Delivery

Jiang

Liu

et al. 2010

Macro Chemistry & Physics

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Reducible disulfide‐containing hyperbranched PEI‐SS‐HP was synthesized via click chemistry and evaluated as nonviral gene carrier. The structure of the polymers was confirmed by 1H NMR and FTIR. It was shown that the PEI‐SS‐HP was able to bind plasmid DNA to positively charged nanoparticles. The reduction sensitivity of the PEI‐SS‐HP was confirmed by gel retardation assay in the presence of DTT mimicking an intracellular reductive environment. In vitro experiments revealed that the reducible PEI‐SS‐HP not only had much lower cytotoxicity, but also posed superior transfection activity as compared to non‐degradable 25‐kDa PEI. The results indicate that a reducibly degradable PEI‐SS‐HP can be a promising gene vector.

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Section: Synthesis Of Pei-ss-hp (7) Via Click Reactionmentioning

confidence: 99%

Disulfide‐Containing Hyperbranched Polyethylenimine Derivatives via Click Chemistry for Nonviral Gene Delivery

Jiang

Liu

et al. 2010

Macro Chemistry & Physics

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“…The interaction between BPEI molecules and the packing material, which disturbs separation efficiency and reproducibility. [9] Salts are often added in the mobile phase to suppress these interactions. But, the addition of salt makes the MW measurement more complex [12].…”

Section: Discussionmentioning

confidence: 99%

“…The MW of polymers can be obtained by using a calibration curve that relates the log of the MW to the retention time or volume [9]. Nowadays, some detectors are employed in tandem with on-line sample fractionation by SEC, especially refractive index (RI) detector and MALLS detector [22][23][24].…”

Section: Size-exclusion Chromatography (Sec) Measurementmentioning

confidence: 99%

“…Besides, High MW BPEI is also more toxic, reducing the viability of cells for transgene expression. In general, the cytotoxicity increases with increasing MW of the cationic polymer [8,9]. There is always a tradeoff between tight PEI/DNA complex formation outside cells and easy dissociation of nucleic acid inside cells, high transfection efficiency and low cytotoxicity, which depends largely on the MW of BPEI.…”

Section: Introductionmentioning

confidence: 99%

“…However, the additional salts can also make it more complex and difficult to the MW measurement [12]. For example, when the salt concentration is too high, hydrophobic interactions may occur [9,11]. Indeed, the actual MW of a given BPEI product appears to be reported differently depending upon the particular company supplying the chemical [2].…”

Section: Introductionmentioning

confidence: 99%

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Colorimetric determination of molecular weight of branched polyethylenimine using unmodified gold nanoparticles

Ling¹

2016

abp

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Branched polyethylenimine (BPEI) is a cationic polymer that have been used to deliver genes and often served as a -golden standard‖ of non-viral vectors. The transfection efficiency/cytotoxicity profile of BPEI depends on its molecular weight (MW). However, due to the strong positive charge of this polyelectrolyte, it is difficult to determine accurately the MW of BPEI, even if applying sophisticated instruments and complicated protocols. Here, we show a colorimetric assay for the measurement of BPEI MW, based on the aggregation of unmodified gold nanoparticles (Au NPs). We found that the different MW of BPEI (1.25 μg/ml) leads to the different aggregation degrees of Au NPs and the color changes of the solutions. Thus, the MW can be distinguished straightforwardly by visual inspection and determined by spectrophotometry. This novel approach is simple (mixing solutions at room temperature), rapid (within 15 min) and sensitive (only depending on the MW).

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The role of polymer nanolayer architecture on the separation performance of anion‐exchange membrane adsorbers: Part II. DNA and virus separations

Bhut

Weaver

Carter

et al. 2011

Biotech & Bioengineering

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The surface-initiated polymerization protocol developed in part I was used to prepare strong anion-exchange membranes with variable polymer chain graft densities and degrees of polymerization for DNA and virus particle separations. A focus of part II was to evaluate the role of polymer nanolayer architecture on DNA and virus binding. Salmon sperm-DNA (SS-DNA) was used as model nucleic acid to measure the dynamic-binding capacities at 10% breakthrough. The dynamic-binding capacity increases linearly with increasing poly ([2-(methacryloyloxy)ethyl]trimethylammonium chloride) chain density up to the highest chain density used in this study. The new membranes yielded threefold higher SS-DNA-binding capacity (30 mg/mL) than a leading commercial membrane with the same functional group chemistry. Elution of bound DNA yielded a sharp peak, and resulted in a 13-fold increase relative to the feed concentration. This concentration effect further demonstrates the highly favorable transport properties of the newly designed Q-type membranes. However, unlike findings in part I on protein binding, SS-DNA binding was not fully reversible. Minute virus of mice (MVM) was used as model virus to evaluate the virus clearance performance of newly designed Q-type membranes. Log reduction of virus (LRV) of MVM increased with increasing polymer chain density. Membranes exhibited >4.5 LRV for the given MVM impurity load and may be capable of higher LRV values, as the MVM concentration in the flow-through fraction of these samples was below the limit of detection of the assay.

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Molar-Mass Characterization of Cationic Polymers for Gene Delivery by Aqueous Size-Exclusion Chromatography

Cited by 63 publications

References 41 publications

Disulfide‐Containing Hyperbranched Polyethylenimine Derivatives via Click Chemistry for Nonviral Gene Delivery

Disulfide‐Containing Hyperbranched Polyethylenimine Derivatives via Click Chemistry for Nonviral Gene Delivery

Colorimetric determination of molecular weight of branched polyethylenimine using unmodified gold nanoparticles

The role of polymer nanolayer architecture on the separation performance of anion‐exchange membrane adsorbers: Part II. DNA and virus separations

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