Influence of Cell Disruption Methods on the Recovery and Immunogenicity of a Fusion Protein for a Therapeutic Cancer Vaccine Against HPV

by Miladys Limonta, Laura Varas, Jorge Valdés, Lourdes Zumalacárregui, Dayana Soler, Maelys Miyares, Alain B. Alfonso, Milaid Granadillo, and Isis Torrens
Volume 14, Issue 4 (Winter 2015/2016)

Two cell disruption methods, mechanical and chemical, were applied for the recovery of a fusion protein named CIGB 550-E7, expressed on Escherichia coli grown in defined saline media. A comparison of the methods was done, and various operating parameters for each technique were optimized to obtain the maximum disruption efficiency and CIGB 550-E7 protein release. The mechanical disruption’s yield and recovery were 1.24 and 1.37 times higher than those obtained with chemical disruption. Modified conditions were assayed for the CIGB 550-E7 obtained by chemically defined media using the mechanical and chemical cell disruption methods. The processes we developed allowed us to obtain an active pharmaceutical ingredient that fulfills the requirements stipulated by the regulatory authorities in terms of purity and lipopolysaccharide contaminants. In addition, the CIGB 550-E7 obtained from both methods showed similar biological activity so that either method could be used. Finally, a cost/benefit relationship (CBR) analysis was done for both disruption methods, and the CBR value for mechanical cell disruption demonstrated that this was the most feasible choice...

Citation:
Limonta M, Varas L, Valdés J, Zumalacárregui L, Soler D, Miyares M, Alfonso AB, Granadillo M, Torrens I. Influence of cell disruption methods on the recovery and immunogenicity of a fusion protein for a therapeutic cancer vaccine against HPV. BioProcess J, 2016; 14(4): 22–9. http://dx.doi.org/10.12665/J144.Limonta.

Posted online January 12, 2016.