Compatability and Stability of Human Serum Albumin and Amino Acid Formulation

Compatibility of amino acid solution in combination with human serum albumin

and effect on measured molecular size.

Introduction:  Serum albumin is an important colloid in the circulation and the interstitial compartments of mammals.  It is an important transporter of proteins as well as endogenous and exogenous materials in the circulation.  Albumin is a potent antioxidant and free radical scavenger and supports endothelial stabilization.1 The use of human serum albumin as a resuscitative fluid has long been established but with mixed reviews as to its superiority over crystalloid solutions.2 

During the sterilization of human serum albumin, the albumin is treated by cold ethanol fractionation followed by ultra- and diafiltration.  The manufacturing process also includes container pasteurization and additional bulk pasteurization.  While this process affords a significant viral reduction in in vitro studies, the combination of process steps including Cohn fractionation and final container pasteurization strip away the proteins and other molecules typically bound to albumin in normal circulation.

Combining human serum albumin and amino acids prior to administration into the circulation has been noted to improve clinical outcomes in the treatment of shock states in unpublished case reports.3 It is postulated that the combination of human serum albumin to amino acids increases the size of the albumin molecule and improves or restores some of the physiological characteristics of albumin before it is administered.  It is also postulated that the combination is compatible and stable and does not precipitate or agglutinate and has good homogeneity.  The purpose of this study is to evaluate compatibility and stability of combining albumin and amino acids at different pH’s and to quantify the size of the molecule created by the combination of albumin and a mixed amino acid solution.

Methods: To measure the “size” of the albumin protein particle in solution we used dynamic light scattering with a laser beam directed into a solution of albumin.  Analysis of the collected data produced a particle size distribution table.  Two types of size distribution tables were generated.  One based on “particle size” and one based on “particle mass”.  Dynamic light scattering data regarding the albumin particle size (assumed as a spherical particle), its apparent molecular weight, and the dispersity of the sample. Dispersity or polydispersity measures were collected to assess the distribution of particles with different sizes (molecular weights) in the sample.  A solution of albumin with little or no aggregation should show low polydisperisity.  Measured polydispersity of albumin protein samples was collected to assess differences in glycosylation between individual albumin molecules, transient formation of dimers, and aggregation in solution.

Measurements were taken from samples of a human serum albumin and amino acid compound at different pH values and concentrations. 

Key Findings:

• The albumin / amino acid formulation is stable at 4°C for at least four weeks and degradation or aggregation does not occur.

• Adding [amino acid formulation] to an albumin solution increases the apparent size of the albumin molecular by up to 17%.

• Polydispersity of 5% albumin / [specific percent specific amino acid] is preferred for clinical application.

THE SPECIFIC FORMULATION CAN BE OBTAINED AND USED ROYALTY FREE FOR A PERIOD OF ONE YEAR BY CONTACTING US HERE.

THE FULL COMPATIBILITY STUDY CAN BE OBTAINED BY CONTACTING US HERE.

REFFERENCES

1.  Horstick G, Lauterbach M, Kempf T, et al. Early albumin infusion improves global and local hemodynamics and reduces inflammatory response in hemorrhagic shock. Crit Care Med. 2002;30(4):851-855.

2.  Quinlan GJ, Mumby S, Martin GS, et al. Albumin influences total plasma antioxidant capacity favorably in patients with acute lung injury. Crit Care Med. 2004;32(3):755-759.

Millam D. The history of intravenous therapy. J Intraven Nurs. 1996;19(1):5-14.3.

3. Millam D. The history of intravenous therapy. J Intraven Nurs. 1996;19(1):5-14.

4.  Foster, Peter (1994). The Kirk -Othmer Encyclopedia of Chemical Technology, 4th edition, vol 11, 990-1021. pp. 990–1021.

5. Cohn, E. J.; Strong, L. E.; Hughes, W. L.; Mulford, D. J.; Ashworth, J. N.; Melin, M.; Taylor, H. L. (1946). "Preparation and Properties of Serum and Plasma Proteins. IV. A System for the Separation into Fractions of the Protein and Lipoprotein Components of Biological Tissues and Fluids 1a,b,c,d". Journal of the American Chemical Society. 68 (3): 459–475.