AIM: To examine body fluids such as ascitic fluid (AF),saliva,bile and pleural effusions for the presence of hepcidin using a novel radioimmunoassay (RIA).METHODS: Serum samples were collected from 25 healthy voluntee...AIM: To examine body fluids such as ascitic fluid (AF),saliva,bile and pleural effusions for the presence of hepcidin using a novel radioimmunoassay (RIA).METHODS: Serum samples were collected from 25 healthy volunteers (mean age: 36 ± 11.9 years,11 males,14 females).In addition bile was obtained from 12 patients undergoing endoscopic retrograde cholangiopancreatography (mean age: 66.9 ± 16.7 years,M:F = 5:7).Saliva was collected from 17 healthy volunteers (mean age: 35 ± 9.9 years,M:F = 8:9).Pleural and AF were collected from 11 and 16 patients [(mean age: 72 ± 20.5 years,M:F = 7:4) and (mean age: 67.32 ± 15.2 years,M:F = 12:4)],respectively.All biological fluid samples (serum,exudative and transudative fluids) were tested for the presence of hepcidin-25 molecule using RIA.RESULTS: Hepcidin-25 was detected in all biological fluids tested.The mean ± SD hepcidin-25 in serum was 15.68 ± 15.7 ng/mL,bile 7.37 ± 7.4 ng/mL,saliva 3.4 ± 2.8 ng/mL,exudative fluid 65.64 ± 96.82 ng/mL and transudative fluid 14.1 ± 17.8 ng/mL.CONCLUSION: We provide clear evidence that hepcidin-25 is present in bile,saliva,pleural and ascitic fluids.Hepcidin is likely to play a role here in innate immunity.展开更多
在大肠杆菌中表达的重组Hepcidin融合蛋白以包涵体形式存在,其N端带有6个组氨酸。以Ni2+-IDA-Sepharose Fast Flow为层析介质,在变性条件下以不同的咪唑和pH值洗脱方式对Hepcidin融合蛋白的纯化效果进行了比较,确定了该融合蛋白的金属...在大肠杆菌中表达的重组Hepcidin融合蛋白以包涵体形式存在,其N端带有6个组氨酸。以Ni2+-IDA-Sepharose Fast Flow为层析介质,在变性条件下以不同的咪唑和pH值洗脱方式对Hepcidin融合蛋白的纯化效果进行了比较,确定了该融合蛋白的金属螯合层析纯化条件。以60 mmol/L咪唑洗脱杂蛋白,然后将pH值降至4.0洗脱融合蛋白,纯化后的融合蛋白纯度大于95%,而且不含咪唑,有利于下一步Hepcidin的制备。金属螯合层析中融合蛋白收率不低于90%。Ni2+-IDA-Sepharose Fast Flow对该融合蛋白的吸附量为30.4 mg/mL。展开更多
基金Supported by Grant from Ealing Hospital NHS Trust,Imperial College,United Kingdom
文摘AIM: To examine body fluids such as ascitic fluid (AF),saliva,bile and pleural effusions for the presence of hepcidin using a novel radioimmunoassay (RIA).METHODS: Serum samples were collected from 25 healthy volunteers (mean age: 36 ± 11.9 years,11 males,14 females).In addition bile was obtained from 12 patients undergoing endoscopic retrograde cholangiopancreatography (mean age: 66.9 ± 16.7 years,M:F = 5:7).Saliva was collected from 17 healthy volunteers (mean age: 35 ± 9.9 years,M:F = 8:9).Pleural and AF were collected from 11 and 16 patients [(mean age: 72 ± 20.5 years,M:F = 7:4) and (mean age: 67.32 ± 15.2 years,M:F = 12:4)],respectively.All biological fluid samples (serum,exudative and transudative fluids) were tested for the presence of hepcidin-25 molecule using RIA.RESULTS: Hepcidin-25 was detected in all biological fluids tested.The mean ± SD hepcidin-25 in serum was 15.68 ± 15.7 ng/mL,bile 7.37 ± 7.4 ng/mL,saliva 3.4 ± 2.8 ng/mL,exudative fluid 65.64 ± 96.82 ng/mL and transudative fluid 14.1 ± 17.8 ng/mL.CONCLUSION: We provide clear evidence that hepcidin-25 is present in bile,saliva,pleural and ascitic fluids.Hepcidin is likely to play a role here in innate immunity.
文摘在大肠杆菌中表达的重组Hepcidin融合蛋白以包涵体形式存在,其N端带有6个组氨酸。以Ni2+-IDA-Sepharose Fast Flow为层析介质,在变性条件下以不同的咪唑和pH值洗脱方式对Hepcidin融合蛋白的纯化效果进行了比较,确定了该融合蛋白的金属螯合层析纯化条件。以60 mmol/L咪唑洗脱杂蛋白,然后将pH值降至4.0洗脱融合蛋白,纯化后的融合蛋白纯度大于95%,而且不含咪唑,有利于下一步Hepcidin的制备。金属螯合层析中融合蛋白收率不低于90%。Ni2+-IDA-Sepharose Fast Flow对该融合蛋白的吸附量为30.4 mg/mL。
