Study On The Preparation Of Human Serum Albumin From Fraction Iv Precipitation

Human serum albumin(HSA)is a plasma soluble protein that is most abundant in human plasma.It is transformed and synthesized in kidney and liver cells.It is essentially a simple plasma monomer soluble protein without any prosthetic groups and Lipids or other carbohydrates bound in the form of covalent carbon-hydrogen bonds.Clinically,human blood albumin has many biological structural properties and types,including combination with organic anions and inorganic cations,ionic coupling with other drugs,drug targeting,antioxidant activity,and enzyme activity.China is the first country in the world to consume human albumin,and domestic products are in short supply.Although China’s blood product industry is developing rapidly,the amount of plasma collected has been steadily increasing every year.However,imported human albumin products are also hitting the domestic market,and the proportion of imported products sales has increased year by year,and has exceeded 60% in 2018.At present,the yield of human blood albumin is 2400-3000 bottles / ton of plasma,and the extraction rate accounts for only 60-70% of the total albumin.During the extraction process,albumin was lost due to various reasons,and the most important part of the loss was the precipitation of Fraction Ⅳ.Therefore,the preparation of human albumin from the precipitation of Fraction Ⅳ has very important theoretical significance and application value.In this paper,Cohn Fraction Ⅳ is used as the raw material,and the low temperature ethanol method is used to prepare albumin in the laboratory.Among them,the dissolution of Fraction Ⅳ dissolved solution,the separation conditions of Fraction Ⅳ-1 and the separation conditions of Fraction Ⅳ-2 were investigated.The effect of p H,ethanol concentration and reaction temperature on the separation of albumin.The screening scheme used(L934)orthogonal test to examine the effect of 3 factors and 3 levels on the yield and quality of albumin.After that,the analysis results and raw data obtained under these separation conditions were subjected to analysis of variance to achieve a reasonable screening of the best separation methods and conditions.In addition,in the ultrafiltration stage,the effect of increasing the sodium chloride concentration to 1.1% on the removal of aluminum ions in albumin products was also investigated.After establishing the parameters of each step,the same batch of Fraction Ⅳ precipitation was used to take a verification scale test of 100.00 Kg.And increase the investigation of the reduction rate of PKA for each separation step.Finally,the 3 batches of finished products were tested and checked for stability.This study found that the human blood albumin yield can reach 160 g / Kg Fraction Ⅳ precipitation,the albumin recovery rate is 65.7%,and the yield is higher;the purity is 97.3 ± 0.1%,which is greater than 96% required by the Pharmacopoeia;the PKA content is reduced Obviously,it is 7.33 ± 1.53 IU / ml,which is far less than the 35 IU / ml required by the pharmacopoeia limit;the aluminum ion content is 21.3 ± 3.79μg / L,and the increase in the storage period is small,and the results of 18 months of stability inspection are all Less than 100μg / L,less than 200μg / L required by the pharmacopoeia limit;polymer 1.43 ± 0.057%,also less than 5% required by the pharmacopoeia limit;the product can withstand the thermal stability test at 57 ℃ 50 h;After the stability inspection,the above main indicators change little and the quality is stable.