Evaluation of Albumin Content in Hyperimmune Plasma and Antivenom for Assessment of Purity

One of the important issues for quality control of biological products is purity. In most cases, electrophoresis is used to determine the purity of the proteins and the quantity of related band is determined by densitometry and respective software. In order to check the purity of hyperimmune plasma and antivenom bulks, we have evaluated the albumin content of these products. Sixty samples were divided into two groups: the hyperimmune plasma group that containing thirty bulks of monovalent and polyvalent anti-snake, polyvalent anti-scorpion and anti-diphtheria toxin plasma, and plasma-derived antivenoms group, comprising thirty bulks sera obtained from the purification of relevant plasma bulks. The albumin purification from horse normal serum was performed through heat shock strategy and used as reference for the quantification of albumin content. After electrophoresis of samples, the gel images were analyzed using Image J software and the amount of protein in relevant band was determined by calculating the curve area and comparison with reference albumin. The results revealed significant changes on albumin level in hyperimmune plasma samples. Two out of 30 plasma samples had lower and sixteen samples had a higher albumin than normal levels. Using the Image J, we could not detect any residue of albumin in antivenom sera. The results demonstrated effectiveness of purification protocol for removal of albumin during antivenom production process. However, development of a quantitative evaluation method along with the semi-quantitative method used in this study can help the accuracy of these results.