International Journal of Biomedical Science and Research
Fast and Cheap Purification Several Significant Blood Plasma Proteins by Casmac: Conclusions from the Two Practical De-velopments
Abstract
Serhiy P. Havryliuk, Olena S. Gavryliuk and Heorhii L. Volkov
Recently we have demonstrated the possibility of using a number of high-affinity peptides to isolate both the total pool of IgG1-4 and specific IgG against several human viruses. Our published list of seven peptides that high specificity interacted with IgG1-4 at seven different points, which was specially made to tightly bind a large immunoglobulin molecule and deprive it mobility in order to avoid denaturation during high-temperature (35-45 °C) virus-inactivation directly in the chromatographic column. The affinity-7-peptide-chromatographic gel was named 7-PEPs-WorkBeads. The affinity peptide for aRUBIgG was developed on the base of interaction of amino acid sequence of RV capsid proteins which are possible the sites of RV interaction with target cell membranes or mitochondrial proteins or RV cell receptor MOG. In a similar way, affinity peptides to IgG against viruses rabies, measles, poliomyelitis and herpes were developed.
Methods: Affinity peptides, designed for chromatographic purification other plasma proteins, also revealed favorable properties for specific capture of Fg, Cpm, ProThr and A1AT family as for high types of buffer flow, as well as high virus inactivation temperature. The study aimed to present highly selective peptides as ligands for separating groups and individual proteins and possibility the “ledges” for the cascade chromatographic separation of human plasma proteins on the basis of the peptide-affinity gels. Human total IgG and subsequent specific anti-virus IgGs (anti-Rubella, anti-Mumps, anti-Measles, anti-Polio and anti-Varicella zoster viruses IgGs) as well as human blood plasma proteins (fibrinogen, ceruloplasmin, prothrombin and alpha-1-antithrombin family) were purified by two specific affinity cascade chromatography methods.
Results: The content of varicella-zoster, mumps, rubella, polio and rabies viruses IgGs were shown corresponded to dates of other experiments with minor deviations. The enough satisfactory purity (>94%) of four plasma proteins and their high yield (>84%) were achieved not only due to the specific affinity of the synthesized gels, but also due to a significant reduction in the buffer volumes, which ensured a significant reduction in the time of oxidation, denaturation and hydrolytic degradation of proteins, that is, the cascade chromatography method used.
Conclusion: As a result, the examples presented in the paper strongly support the transformation of biologics purification towards multi-column chromatography instrumentation due to its beneficial effects on product quality, time savings, and reduction of production costs. Moreover, these methods can be easily scaled to industrial processes.