Response surface methodology (RSM) is used to optimize the medium of Tetraselmis sp.-1 which is cell fused microalgae capable of growing under mixotrophic condition. Empirical models are developed to describe the rela...Response surface methodology (RSM) is used to optimize the medium of Tetraselmis sp.-1 which is cell fused microalgae capable of growing under mixotrophic condition. Empirical models are developed to describe the relationships between the operating variables (glucose, urea, sodium dehydrogenate phosphate, sodium chloride) and responses (cell density). Statistical analysis indicates that glucose and urea have significant effects on the microalgae cell density, but other two factors (sodium dehydrogenate phosphate, sodium chloride) have no obvious effect. The path of steepest ascent is used to approach the optimal region of medium composition. Optimal cell density (2.638 g dry weight/L) was reached when the operating conditions were glucose concentration (30.75 %), urea concentration (0.440 g/L), sodium dehydrogenate phosphate (15 mg/L) and sodium chloride (28 g/L).展开更多
Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency....Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.展开更多
文摘Response surface methodology (RSM) is used to optimize the medium of Tetraselmis sp.-1 which is cell fused microalgae capable of growing under mixotrophic condition. Empirical models are developed to describe the relationships between the operating variables (glucose, urea, sodium dehydrogenate phosphate, sodium chloride) and responses (cell density). Statistical analysis indicates that glucose and urea have significant effects on the microalgae cell density, but other two factors (sodium dehydrogenate phosphate, sodium chloride) have no obvious effect. The path of steepest ascent is used to approach the optimal region of medium composition. Optimal cell density (2.638 g dry weight/L) was reached when the operating conditions were glucose concentration (30.75 %), urea concentration (0.440 g/L), sodium dehydrogenate phosphate (15 mg/L) and sodium chloride (28 g/L).
基金supported by the National Basic Research Program of China (2011CB915502)State Key Laboratory of Proteomics Project (SKLP Y200907)National High Technology Research and Development Program of China (2012AA020201)
文摘Recognizing proteins via the production of highly specific monoclonal antibodies(mAbs)is crucial to identifying proteins for proteomic research.However,traditional mAb generation is time-consuming with low efficiency.In this study,we assessed the high throughput method of producing mAbs by immunizing mice with multiple antigens in order to obtain hybridomas against these multiple antigens in one cell fusion.We selected eight proteins that play important roles in human physiological or pathological processes.These proteins were mixed and simultaneously administered to one mouse.We observed the immunizing period for 10 d,and determined the effect of liquid medium and semi-solid medium in hybridoma generation.As a result,all eight immunogens induced antibodies in the immunized mouse in one cell fusion,we obtained hybridomas specific to all eight proteins by enzyme-linked immuno sorbent assay(ELISA)screening,hybridomas against five out of eight showed specific positive in Western-blotting assays.This indicates that we generated mAbs specific to eight proteins in one cell fusion,greatly increasing the efficiency of mAb generation.Furthermore,we observed that hybridomas selected from the liquid medium and semi-solid medium showed different reactivity to antigens.Our study established high-throughput and time-saving methods for production of mAbs.These results provide alternative approaches for increasing the efficacy of mAb generation.
