アルブミン (ヒト由来, フラクションV) 化学特性,用途語,生産方法
外観
白色～黄褐色、結晶～粉末又は塊
溶解性
水に溶け、エタノールに難溶である。
解説
DNAとハイブリダイゼーションを起こさせることで，特定のmRNAを探索する手法．DNA-DNAハイブリダイゼーションをサザンブロット法とよぶことに由来してつけられた．
使用上の注意
アルゴン封入
説明
Albumin is a known carrier of fatty acids (FA). Thus control over specific FA′s for cell culture is important, as different cell lines can differ in their sensitivity to particular fatty acids. Fatty acid-free human serum albumin (HSA) is therefore useful for cell culture studies where specific fatty acid content must be strictly controlled, so that researchers can use particular fatty acids specific to their cell lines. Fatty acid-free albumin also allows for optimal and maximum binding sites for using specific fatty acids in cell culture. The use of FA-free HSA also addresses concerns about endogenous FA′s potentially in non-FA-free HSA.
化学的特性
solid
使用
Albumin from human serum has been used in the preparation of glycated human serum albumin (gHSA) in a glucose solution. It has also been used in the study to estimate the impact of surface nanotopography and chemical composition on blood compatibility.
主な応用
Albumin from human serum has been used in the preparation of glycated human serum albumin (gHSA) in a glucose solution.
It has also been used in the study to estimate the impact of surface
nanotopography and chemical composition on blood compatibility.
Albumin was used to test its effect on the in vitro bactericidal activity of cefditoren against penicillin-resistant Streptococcus pneumonia.
It has been clinically used in serious and often life-threatening
conditions, such as shock and blood loss due to trauma, burns, and
surgery. It was used also to test the effect of non-enzymatic glycation on the unfolding of human serum albumin.
一般的な説明
Prepared under non-denaturing conditions by a modification of the Cohn procedure.
合成
The synthesis of human serum albumin (HSA) comprises both in vivo and in vitro pathways. In vivo, HSA is synthesized exclusively by the liver: its mRNA encodes a 609-amino-acid precursor, pre-pro-albumin, which enters the endoplasmic reticulum, where the N-terminal 18-amino-acid signal peptide is rapidly cleaved, followed by removal of the N-terminal basic 6-amino-acid pro-peptide just before secretion, yielding the mature non-glycosylated monomer of 585 amino acids with a molecular mass of about 66.5 kDa; a healthy adult liver synthesizes approximately 13.8 g of albumin per day, accounting for about 25% of total hepatic protein synthesis[1].
In vitro, recombinant HSA was produced in E. coli by cloning the full coding sequence of mature HSA from a human liver cDNA library into expression plasmids such as pXL276 and its derivatives (with the PL promoter replacing the trp promoter); the recombinant protein was mainly produced as insoluble aggregates, which were resolubilized, refolded by 100-fold slow dilution into refolding buffer, and purified by anion-exchange FPLC with an average final yield of about 40% at laboratory scale[2]. To obtain an authentic N-terminus, a fusion strategy was adopted in which the first six residues of phage λcII protein were fused to mature HSA to boost expression; after refolding, the fusion protein was cleaved in vitro with catalytic amounts of trypsin (<1/1000 w/w), releasing a mature recombinant HSA whose unique N-terminal sequence matched native HSA, and the product was indistinguishable from native HSA by amino acid composition, UV absorption spectroscopy, electrophoretic and chromatographic analyses.
アルブミン (ヒト由来, フラクションV) 上流と下流の製品情報
原材料
準備製品