Gradient pH co-precipitation to recover proteins from plasma discarded in artificial-liver treatment
- VernacularTitle:梯度pH共沉淀法回收人工肝丢弃血浆蛋白初步实验
- Author:
Bing XU
;
Chen PAN
;
Xiao-Zhi JIANG
;
Man-Zhi LIN
;
Min CHEN
;
Zhuo LIN
;
Yi CHEN
;
Hui-Cong CHEN
;
- Publication Type:Journal Article
- Keywords:
Liver,artificial;
Hydroge-ion concentration;
Precipitation;
Salting out;
Blood proteins;
Recovery
- From:
Chinese Journal of Infectious Diseases
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To set up a system in vitro to rapidly recover plasma proteins lost during artificial-liver treatment.Methods The polyprotein precipitation was obtained by all proteins whose isoelectric point pH value were between 7.3 and 5.1,which collided with each other and aggregated using gradient pH co-precipitation(adding 1 mol/L citric acid slowly in the plasma solution to change the pH values gradually from 7.3 to 5.1 in 5 h)combined with salting out(degree of saturation of NaCl is 33%,reacted for 5.5 h at 4℃)or low-temperature ethanol precipitation(40% ethanol, reacted for 5.5 h at -7℃)so that to get rid of toxicants by discarding the supernatant.Results In the range of pH 5.1-7.3,50%(29g/57g)of the total plasma proteins had been recovered by the gradient pH salting out and 41%(25 g/61g)by the gradient pH low-temperature ethanol co-precipi- tation.The protein remained in the supernatant was mostly albumin and its combined bilirubin.The levels of total bilirubin decreased to 0.07% and 0.06% of the original levels by these two methods respectively and the serum HBV DNA level decreased to be undetected(quantitative PCR).Conclu- sions The proteins with close isoelectric point can co-precipitated with the presence of high concen- tration of NaCl or low-temperature ethanol and by changing the pH value gradually.The total protein in the discarded plasma during artificial-liver treatment can be recovered rapidly using the gradient pH coprecipitation.
