Abstract:

Abstract:Objective To establish  the method for detection of biological activity of basic fibroblast growth factor(bFGF )protein sponge in vitro and investigate the stability of bFGF protein sponge stored  at room temperature.Methods The bFGF biological protein sponge was fused into the dilution of the test solution,the extract was obtained after centrifugation,the bFGF standard samples were used as   control,and the bFGF protein sponge was cultivated with NIH3T3 cells.MTT assay was used to dected the proflieration  of NIH3T3 cells induced with bFGF biological protein sponge extract.The   bFGF protein  sponge stored for 2,3,4 years at room temperature was used as  the test materials,the bFGF biological protein sponge stored for 30 d at low temperature  as  standard  control,the culture medium without  bFGF protein  sponge and  bFGF  as blank control,the  potency of biological activity of each group was detected.The   stability of biological activities in vitro  of  bFGF protein sponge stored for different time were compared.Results Compared with blank  control,the A value of bFGF biological protein sponge extract was increased with the increasing of concentration,in other words the  NIH3T3 cell proliferation was increased(P<0.01);compared with bFGF standard control, the difference was not significant (P> 0.05),either bFGF standard or bFGF biological protein sponge showed a dose-response relationship  by the developed method,and the determination result complied with the following four-parameter equation:y =（A-D）／［1 +（X ／ C）B］+D. Compared with protein sponge stored for 30 d at low temperature,the descending rate of biological activity titers (U?mL-1) of the protein sponge stored for 2 to 4 years at room temperature  were  0.5%,0.6% and 0.8%,lower than the national standard of 15%. Conclusion The NIH3T3 cell proliferation could be applied for detection of the biological activity of bFGF protein sponge in vitro,it could be used as the routine detection method for bFGF protein sponge,the stabilities of bFGF protein sponge stored for 2-4 years at room temperature are good.

Key words: basic fibroblast growth factor, biologic protein sponge;biological activitydrug