ABSTRACT
Objective To screen out the bioactive sulfate saccharides interacted with granulocyte colony-stimulating factor (G-CSF) and provide some scientific data on their pharmacology. Methods The interactions between G-CSF and 15 sulfate saccharides with various chain lengths and extent of sulfation were studied by capillary electrophoresis. Furthermore, the effects of these G-CSF binding saccharides on the proliferation and differentiation of leukemia cell line and NFS-60 were also investigated. Results Except for the hepatosaccharide, degradable carrageenan, Dextran sulfate (DexS) with a low molecular mass the nonsulfated sancharide and alkaline chitosan oligosaccharide, the rest of saccharides had interact with G-CSF.DexS500, λ carrageenan, ι carrageenan, κ carrageenan and low molecular weight heparin-1(LMWH-1)with anticoagulant activity could inhibit the NFS-60 growth. Compared with the absorbency of the control group (100.00±3.00) after these saccharides interacted with the NPS-60 cells, their relative absorbencies were 31.33±2.08,37.67±2.08,45.00±1.20,43.00±2.65 and 74.67±1.52, respectively. Their differences had a statistical significance(t=27.94,26.71,26.42,26.61,10.54,P<0.01). The morphological analytic results showed that the LMWH and lambda carrageenan as well as DexS with a molecular mass of 500 000 induce effectively promyclocyte to transit metamyelocyte. Flow cytometry results showed enhanced expressions of CD11b, CD18 and CD45 after treatment with the LMWH, lambda carrageenan and DexS500,respectively. Cell cycle analysis indicated that the S phase population decreased after treatment with these saccharides. Conclusions Capillary electrophoresis is an effective medicine-screening technique that is used to screen some of these saccharides which could bind to G-CSF. And it is found that the interactions are correlated with the chain length, the extent and the position of sulfation of these saccharides.
ABSTRACT
In this review, the analytical methods of polyethylene glycol-modified proteins are introduced, including spectroscopy, liquid chromatography, electrophoresis, mass spectrometry, etc. These methods are compared in respect to their individual characteristics.
ABSTRACT
Glycerol,mannitol has been used as additives for the separation of deoxyribonucleic acid (DNA)fragments in entangled solution by capillary electrophoresis. Differing from glycerol and mannitol,the structure of glucose is a ring with 5 hydroxyl groups.This structure helps it easily to form complex with boric acid and hydroxypropyl methylcellulose (HPMC).On the other hand,glucose has large space obstacle compared with glycerol and mannitol.So glucose can markedly improve separation of DNA in HPMC solution by capillary electrophoresis compared with other additives.The factors such as boric acid concentration and pH,influencing formation of complex of HPMC and glucose were studied. High concentration of boric acid could improve the DNA separation in glucose-HPMC matrix system. The optimization pH to separate DNA in HPMC-glucose solution by capillary electrophoresis was 8.3.