The percentage of probes captured in the precipitate of the PEI-LAMP product complex was calculated according to the following formula based on the obtained results.
PEI-1, PEI-2, PEI-3, and PEI-4 represent PEI contents of 10%, 40 %, 70, and 100% (w/w totandlipid) in liposomal formulations, respectively.
To remove unbound PEI, the reaction product was washed with ddH2O and centrifuged repeatedly at 3,000 rpm for 15 30 min in an Amicon Ultra-15 Centrifugal Filter Unit (Millipore, Billerica, MA, USA) with a molecular weight cut-off of 100 kDa.
Insolubilization of LAMP products by PEI was inhibited by addition of an excessive amount of KCl to the LAMP reaction solution after amplification.
The relative polyplex ion abundances in the ESI mass spectra reveal the following order of solution stabilities for the major product PN: PEI-d TTTTT) > PEI-d TTTTT) ≈ PEI-d(GCGAT) ≈ PEI-d(CCCCC) > PEI-d GGGGG).
Thus, the guanidinated products G-PEI-P(Orn)n contain multiple poly ornithine-co-arginine) arms.
A solution was prepared by serially diluting a LAMP reaction solution with a known concentration (28 μg DNA/25 μl) using LAMP reaction buffer in order to systematically investigate the reaction between the LAMP product and PEI.
An excess of diethylether was used to precipitate and then wash the product PEI-LA.
The potential environmental impact (PEI) is 44.99 PEI/t of products, with the potential to reduce CO2 emissions by 39% compared with Scenario 1.
We also discuss use of the available pancrelipase products for treating PEI caused predominantly by pancreatic diseases.
