Modification of liposomes with proteins by dansyl-labeled heterobifunctional crosslinker.
The introduction of a fluorescent chromaphore into bifunctional crosslinkers results in a molecule with normal crosslinker properties and a fluorescent group for straightforward quantification. This work describes the synthesis of the dansyl-labeled heterobifunctional crosslinker N-succinimidyl ε-N-dansyl α-N-(acetylthio)acetyllysine (dansyl-ATA-lysine-NHS) containing reactive N-hydroxysuccinimidyl (NHS) ester and sulfhydryl groups. The application of this crosslinker to conjugation of bovine serum albumin (BSA) protein to the surface of a liposome containing maleimide functions is also demonstrated. BSA was modified with the dansyl-labeled crosslinker and subsequently conjugated to liposomes containing reactive phospholipid derivative N-[4-(p-maleimidophenyl)butyryl]phosphatidylethanolamine and the degree of modification and conjugation were quantitatively determined by measuring the fluorescence emission of the dansyl group. The reliability of the fluorescence quantification was confirmed by a micro bio-barcode assay protein assay.
Faculty of Life Sciences, Northwestern Polytechnical University Xi'an, Shaanxi 710072, PR China. taochen@libang.com.cn
Tao
T
Chen
Rutao
R
Wang
Tingting
T
Lu
Guozheng
G
Liang
Tingli
T
Lu
D000818:Animals; D002417:Cattle; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D003619:Dansyl Compounds / Q000138:chemical synthesis / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry*; D008239:Lysine / Q000031:analogs & derivatives / Q000138:chemical synthesis; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013388:Succinimides / Q000138:chemical synthesis / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000138:chemical synthesis / Q000737:chemistry*
D003432:Cross-Linking Reagents; D003619:Dansyl Compounds; D008081:Liposomes; D013388:Succinimides; D013438:Sulfhydryl Compounds; D012710:Serum Albumin, Bovine; D008239:Lysine