Polysciences聚乙烯亚胺（PEI）--转染试剂线性PEI MAX溶液的配制

线性聚乙烯亚胺盐酸盐(Linear Polyethylenimine Hydrochloride)是线性化聚乙烯亚胺PEI 25,000的升级产品，是一种功能更为强大的高效瞬时转染试剂。与PEI 25000的区别在于：1）*水解（去乙酰化）2）PEI 25000的盐酸盐形式，具更高的水溶特性 3）含更长连续性乙烯亚胺片段，其质子化氮水平比PEI25000提高11%以上。

另PEI MAX 40K比PEI 25K更易于使用，并且更高效。PEI 25K转染溶液通常需要几个小时才能制备，而PEI MAX 40K可以在两个小时内转化为即用型溶液。而且PEI 25K含有4-11%的残余丙酰基，可防止聚合物主链与DNA强烈结合。PEI MAX 40K*去乙酰化的结构意味着每批产品的表现始终如一。

下面将主要介绍Polysciences聚乙烯亚胺（PEI） 线性PEI MAX(#24765)的产品特性及其溶液的配制过程。

1.产品特性:

产品名称：转染级线性聚乙烯亚胺盐酸盐(PEI MAX)(Linear Polyethylenimine Hydrochloride)

2.溶液配制

此指导说明书是经polysciences公司研究，测试及其客户反馈的后编写的。下面推荐一种简单适用而高效的方法，讲述如何配制浓度为1mg/mL的转染试剂的配制。

材料：

1g #24765、1L 纯水（Milli-Q®纯水，注射用水（WFI），或类似的生物级水）、1N氢氧化钠、25mL无菌塑料吸管、一次性0.1µm, 0.2µm, 0.22µm PES真空无菌过滤器、无菌高聚乙烯或聚丙烯试剂瓶。

器材：

1 L 玻璃烧杯、1 L玻璃量筒、pH计、移液器、磁力搅拌棒、带油管的真空泵

配制过程：

1g #24765——900mL水溶解——搅拌、加热（60-80°C）——加1N NaOH调pH到6.9-7.1——移液到量筒，加水补足至1L——真空过滤消毒——分装，4°C保存。

备注：加热或加酸都有助于PEI溶解。#24765比#23966易溶，可适当少量加酸，详细用量可试加一点搅拌看看，足够溶解就行，不一定需要加到pH那么低。

PEI MAX - Transfection Grade Linear Polyethylenimine Hydrochloride (MW 40,000)

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DESCRIPTION

PEI MAX 40K (also known as PEI 22K in free base) is a powerful, trusted, and cost-effective transient transfection reagent. In HEK293 and CHO expression systems, PEI offers consistently high gene expression on a wide scale (96 well plates up to 100 L bioreactors). Each year, more researchers and companies turn to Polysciences PEI to gain an edge in their critical work. Relative to most other options, using PEI to prepare transfection reagents in-house can offer as much as a 40% reduction in total transfection costs.

PEI MAX 40K is easier to use and offers consistently higher titers than PEI 25K. PEI 25K transfection solutions typically take several hours to prepare, while PEI MAX 40K can be converted to a ready-to-use solution in under two hours. Additionally, PEI 25K contains 4-11% residual propionyl groups, which prevents the polymer backbone from strongly binding to DNA. PEI MAX 40K’s fully depropionylated structure means each batch performs consistently higher.

Comparison of PEI 25K (#23966) and PEI MAX 40K (#24765).
Method: 10e6/mL HEK293 cells in 50 mL FS transfected with IgG64 plasmid pair. PEI:DNA 4:1. Samples taken 120 hpt. Quantified with Thermo Fisher #23310. N=4 each. Error bar = standard deviation.

A pre-made sterile-filtered solution of PEI MAX 40K is available as our Transporter 5™ Transfection Reagent.

CAS#:

49553-93-7

Molecular Weight:

40,000 (~22,000 free base)

Soluble In:

Cold and room temperature water
Insoluble in: Common organic solvents (ethanol, acetone, tetrahydrofuran)

Appearance:

White to off-white free flowing solid

Reference(s):

Baranyi, L. et al. Rapid Generation of Stable Cell Lines Expressing High Levels of Erythropoietin, Factor VIII, and an Antihuman CD20 Antibody Using Lentiviral Vectors. Human Gene Therapy Methods 24, 214–227 (2013). doi:10.1089/hgtb.2013.002

** Delafosse, L., Xu, P. & Durocher, Y. Comparative study of polyethylenimines for transient gene expression in mammalian HEK293 and CHO cells. Journal of Biotechnology 227, 103–111 (2016). doi:10.1016/j.jbiotec.2016.04.028

** Gutiérrez-Granados, S., Cervera, L., Segura, M. de las M., Wölfel, J. & Gòdia, F. Optimized production of HIV-1 virus-like particles by transient transfection in CAP-T cells. Applied Microbiology and Biotechnology 100, 3935–3947 (2016). doi:10.1007/s00253-015-7213-x

Kobayashi, S., Yoshii, K., Hirano, M., Muto, M. & Kariwa, H. A novel reverse genetics system for production of infectious West Nile virus using homologous recombination in mammalian cells. Journal of Virological Methods 240, 14–20 (2017). doi:10.1016/j.jviromet.2016.11.006

** Longo, P. a, Kavran, J. M., Kim, M. & Leahy, D. J. Transient Mammalian Cell Transfection wtih Polyethylenimine (PEI). Methods Enzymology 529, 227–240 (2013). doi:10.1016/B978-0-12-418687-3.00018-5.

Mann JF, McKay PF, Arokiasamy S, Patel RK, Klein K, Shattock RJ. (2013). Pulmonary delivery of DNA vaccine constructs using deacylated PEI elicits immune responses and protects against viral challenge infection. J Control Release. 170(3):452-9.

** Stuible, M. et al. Optimization of a high-cell-density polyethylenimine transfection method for rapid protein production in CHO-EBNA1 cells. Journal of Biotechnology 281, 39–47 (2018). doi:10.1016/j.jbiotec.2018.06.307

Thomas M, Lu JJ, Ge Q, Zhang C, Chen J, Klibanov AM. (2005). Full deacylation of polyethylenimine dramatically boosts its gene delivery efficiency and specificity to mouse lung. Proc Natl Acad Sci U S A. 102(16):5679-84.

** Contains particularly useful information.

TSCA

Hazards:

Irritant

Handling:

Glove, chemical goggles & fume hood

Storage:

Store at room temperature

MSDS / TECHNICAL DATA SHEETS / PRODUCT LITERATURE

• MSDSMaterial Safety Datasheet 24765 (PDF)

• DATAFT-IR Spectrum

• DATAExample COA

• LITBSE/TSE Statement

• LITTransfection Reagent Preparation and Storage Recommendations

RELATED PRODUCTS

1. Transporter 5™ Transfection Reagent

2. Polyethylenimine, Linear, MW 25000, Transfection Grade (PEI 25K)