Polysciences聚乙烯亞胺（PEI）--轉(zhuǎn)染試劑線性PEI MAX溶液的配制

線性聚乙烯亞胺鹽酸鹽(Linear Polyethylenimine Hydrochloride)是線性化聚乙烯亞胺PEI 25,000的升級(jí)產(chǎn)品，是一種功能更為強(qiáng)大的高效瞬時(shí)轉(zhuǎn)染試劑。與PEI 25000的區(qū)別在于：1）*水解（去乙酰化）2）PEI 25000的鹽酸鹽形式，具更高的水溶特性 3）含更長(zhǎng)連續(xù)性乙烯亞胺片段，其質(zhì)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轉(zhuǎn)染溶液通常需要幾個(gè)小時(shí)才能制備，而PEI MAX 40K可以在兩個(gè)小時(shí)內(nèi)轉(zhuǎn)化為即用型溶液。而且PEI 25K含有4-11%的殘余丙酰基，可防止聚合物主鏈與DNA強(qiáng)烈結(jié)合。PEI MAX 40K*去乙酰化的結(jié)構(gòu)意味著每批產(chǎn)品的表現(xiàn)始終如一。

下面將主要介紹Polysciences聚乙烯亞胺（PEI） 線性PEI MAX(#24765)的產(chǎn)品特性及其溶液的配制過程。

1.產(chǎn)品特性:

產(chǎn)品名稱：轉(zhuǎn)染級(jí)線性聚乙烯亞胺鹽酸鹽(PEI MAX)(Linear Polyethylenimine Hydrochloride)

2.溶液配制

此指導(dǎo)說明書是經(jīng)polysciences公司研究，測(cè)試及其客戶反饋的后編寫的。下面推薦一種簡(jiǎn)單適用而高效的方法，講述如何配制濃度為1mg/mL的轉(zhuǎn)染試劑的配制。

材料：

1g #24765、1L 純水（Milli-Q®純水，注射用水（WFI），或類似的生物級(jí)水）、1N氫氧化鈉、25mL無菌塑料吸管、一次性0.1µm, 0.2µm, 0.22µm PES真空無菌過濾器、無菌高聚乙烯或聚丙烯試劑瓶。

器材：

1 L 玻璃燒杯、1 L玻璃量筒、pH計(jì)、移液器、磁力攪拌棒、帶油管的真空泵

配制過程：

1g #24765——900mL水溶解——攪拌、加熱（60-80°C）——加1N NaOH調(diào)pH到6.9-7.1——移液到量筒，加水補(bǔ)足至1L——真空過濾消毒——分裝，4°C保存。

備注：加熱或加酸都有助于PEI溶解。#24765比#23966易溶，可適當(dāng)少量加酸，詳細(xì)用量可試加一點(diǎn)攪拌看看，足夠溶解就行，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., W?lfel, 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)