›› 2009, Vol. 52 ›› Issue (6): 624-630.doi:

• RESEARCH PAPERS • Previous Articles     Next Articles

Optimization of the method for solid phase extraction of antiviral peptides from haemolymph of diapausing larvae of Musca domestica with methanol.


  • Online:2009-06-20 Published:2009-06-20

Abstract: In order to optimize the conditions of antiviral peptide extraction from haemolymph of diapausing housefly (Musca domestica) larvae with methanol, we compared the separating results of methanol and methyl cyanide and selected the best extraction gradient. After studying the proper protein sample concentration, eluting times, eluting velocity and eluting temperature, we got a better protocol which leads to perfect results of antiviral peptide extraction. Tricine-SDS-PAGE results showed that continuous gradient methanol as an extractive organic solvent eluted more active fractions than methyl cyanide (P<0.05), and the continuous gradient elution with mthanol (10%-100%) was better than interrupted gradient (10%, 30%, 50%, 70% and 90%;20%, 40%, 60%, 80% and 100%, respectively)in the recovery rate and reproducibility of active fractions. Under 0-4℃, when 1.5±0.4 mg/mL haemolymph extracts was added into Sep-Pak C18 catridge and eluted 4 times with the flowing velocity of 1 mL/min, the antiviral fractions were separated better. Upon electrophoresis, only 2 target protein bands of active fractions were eluted by methyl cyanide, while more bands were eluted by methanol.

Key words: Musca domestica, diapause, hemolymph, antiviral peptide, solid phase extract, methanol