Purification of IgG and albumin from human plasma by aqueous two phase system fractionation

Abstract

The current shortages in human plasma products at global levels justify the development of new, cost effective plasma fractionation methods. We have developed a fractionation process to obtain immunoglobulin G (IgG) and albumin-enriched fractions based on polymer-salt aqueous two phase system (ATPS). A small-scale (0.02 L) ATPS composed of polyethyleneglycol 3350 (PEG), potassium phosphate and sodium chloride, at pH 6.1, was evaluated and subjected to 50-fold scale-up (1 L). Further purification of the fractions was performed using caprylic acid precipitation and ion exchange chromatography. Similar yield and purity were obtained at both small and large scales. IgG precipitated in the PEG rich upper phase at 83% recovery and 2.75-fold purification factor. An 81% pure albumin fraction was obtained in the salt rich bottom phase with a 91% yield. After polishing, IgG was obtained at a recovery of 70% and a purity of 92%. Corresponding values for albumin were 91% and 90%. This IgG and albumin fractionation technology deserves further evaluation as it may represent a potential alternative to conventional plasma fractionation methods.

Purification of equine whole IgG snake antivenom by using an aqueous two phase system as a primary purification step

“…Compared to the results presented by Vargas et al [21] for human plasma, a similar behavior was observed between fractionation of human and equine plasma in ATPS. Fig.…”

“…A modification of the procedure for IgG purification from human plasma described by Vargas et al [21] and by the patent application document WO2012136172A1 [24] was employed. A diagram of the methodology is presented in Fig.…”

“…More recently, the primary recovery of human IgG in ATPS was reported when fractionating human plasma for the simultaneous recovery of IgG and albumin in PEG/potassium phosphate ATPS [21]. This approach renders an IgG-enriched upper phase and an albumin-enriched bottom phase, both of which can be further purified.…”

Desenvolvimento de duas estratégias de purificação de IgM a partir de plasma humano.

“…Em alguns casos, a presença de IgM é citada nos processos de produção descritos para IgG VARGAS et al, 2012;VARGAS et al, 2015;WU et al, 2014 Neste trabalho nós propomos utilizar as mesmas técnicas utilizadas por Jehanli e Hough, uma gel filtração seguida de uma troca aniônica. Entretanto, a coluna de gel filtração utilizada neste trabalho, a Sepharose 4 FF, é para uso industrial e podemos aplicar de 20 a 25% volumes de coluna de amostra.…”

“…Alguns trabalhos descritos na literatura visam a purificação de IgG e albumina (FOURCART et al, 1982;GRELLET et al, 2001;VARGAS et al, 2012), mas não há nenhuma descrição conhecida com foco na purificação de IgM. Também é conhecida da literatura a separação de IgG de IgM (BREEN et al, 2012;BRNE et al, 2007;WU et al, 2014).…”