A Simple Method for the Preparation of Human Serum Albumin

Abstract

A simple method, based on ethanol fractionation, for the preparation of highly purified human serum albumin with a higher yield than that of the conventional ethanol procedures is described. It consists of two purification steps, namely, precipitation of most of the other plasma proteins from a 3-fold diluted plasma with ethanol at 42% concentration, pH 5.75 and -5°C, leaving over 96% pure albumin in the supernatant, followed by isolectric precipitation of albumin from the supernatant at pH 4.8 and -5°C. The paste thus obtained was processed to the final albumin solution according to the conventional methods. The yield of the final albumin with a purity of over 99% was equivalent to 29.5 g/l of plasma representing a recovery of over 93%. The possibility of recovering other plasma proteins and the suitability for large scale preparation are also discussed.

Effective removal of albumin from serum

“…Serum protein fractions were obtained using the DAC protocol, which was modified from a previously published method for purifying albumin [7] (Fig. 1).…”

“…Proteins were resolved in the second dimension Figure 1. Schematic of the DAC protocol: (modified from [7]). An amount of 100-500 mL of serum was used.…”

Albumin-Coated Polycaprolactone (PCL)–Decellularized Extracellular Matrix (dECM) Scaffold for Bone Regeneration

“…An additional potent source of calcium-binding and growth-promoting factors that complements NCPs may be serum albumin . Unlike NCPs, albumin is the most abundant protein in the plasma and can be precipitated with high yield and purity . A series of experiments with serum albumin coated on allografts supported mesenchymal stem cell proliferation , and showed improved bone formation in a femoral defect in a rat model …”

“…21 Unlike NCPs, albumin is the most abundant protein in the plasma and can be precipitated with high yield and purity. 22 A series of experiments with serum albumin coated on allografts supported mesenchymal stem cell proliferation 23,24 and showed improved bone formation in a femoral defect in a rat model. 25 This work aims to synergistically combine chondrocytederived decellularized ECM (dECM) and adsorbed serum albumin as potent sources of non-collagenous calcium-binding proteins and evaluate their capacity in tandem to promote osteoblast mineral deposition in vitro.…”

A materials science framework for connecting protein stability with surface adsorption and coating applications

“…Some companies today like Sigma-Aldrich, 217 Gemini Bio-Products, 218 and Equitech-Bio 219 still sell serum albumin fractionated based on Cohn's cold ethanol fractionation method although certain steps are likely to have been modified. [220][221][222] Another fractionation method reported in companies like Sigma-Aldrich, 217 Rocky Mountain Biologicals, 223 Bovogen, 224 Gemini Bio-Products, 218 and Equitech-Bio 225 is heat-shock fractionation. The earliest found record describing a method to purify serum albumin from plasma by heating is in a patent by Reid dated 1955.…”