Capturing of acidic macromolecules from biological samples using a temperature-responsive polymer modified with poly-l-lysine.
We synthesized a temperature-responsive polymer, N-(isopropylacrylamide)-methacrylic acid copolymer, to which poly-l-lysine was introduced. The synthesized polymer as well as the parent polymer showed reversible soluble-insoluble changes in response to temperature changes across the lower critical solution temperature at 32 degree C in an aqueous solution. We found that the polymer efficiently captured acidic bio-macromolecules such as RNA, glycosaminoglycans and mucin-type glycoproteins in biological samples, and the captured molecules were recovered using aqueous NaCl solutions at high concentration. The target acidic molecules thus obtained will be employed for further studies such as structural analysis after brief desalting procedure. The proposed method does not require any chromatographic separations, but only needs a small volume of an aqueous salt solution for releasing captured molecules. Overall procedures are quite easy and simple, and are completed at least within 1 h. We show a few examples for capturing RNA and glycosaminoglycans from cultured cells using the polymer.
Hayashi
Tomonori
T
Faculty of Pharmaceutical Sciences, Kinki University, Kowakae 3-4-1, Higashi-Osaka, 577-8502, Japan.
Yasueda
Shin-Ichi
S
Nakanishi
Yasuharu
Y
Ohta
Hiroko
H
Kinoshita
Mitsuhiro
M
Miki
Yasuyoshi
Y
Masuko
Takashi
T
Kakehi
Kazuaki
K
D000178:Acrylamides / Q000138:chemical synthesis; D000818:Animals; D002417:Cattle; D002477:Cells / Q000737:chemistry*; D002478:Cells, Cultured; D006025:Glycosaminoglycans / Q000032:analysis*; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009077:Mucins / Q000032:analysis*; D011107:Polylysine; D011108:Polymers; D011109:Polymethacrylic Acids / Q000138:chemical synthesis; D012313:RNA / Q000032:analysis*; D013696:Temperature
D000178:Acrylamides; D006025:Glycosaminoglycans; D046911:Macromolecular Substances; D009077:Mucins; D011108:Polymers; D011109:Polymethacrylic Acids; C085085:poly(N-isopropylacrylamide-co-methacrylic acid); D011107:Polylysine; D012313:RNA