FEEDING STRATEGIES AND PURIFICATION PROCESSES FOR MONOCLONAL ANTIBODY PRODUCTION

摘要

This invention provides an improved process for manufacturing a Rabies monoclonal antibody (HuMab 17C7) that results in low osmolality, minimum secondary metabolites like ammonia and lactate, enhanced cell growth and productivity, minimum aggregation or degradation of monoclonal antibody during purification, thereby improving potency of monoclonal antibody (HuMab 17C7) as compared to human rabies immunoglobulin (hRIG).

主权项

1. A method for producing a monoclonal antibody with enhanced potency comprising:
a) adding “Feed solution A” containing vitamins, amino acids and glucose at log phase at a concentration between 0.2% to 0.8% with a flow rate of about 5-15 ml/min at an initial osmolality of about 350 mOSm/kg; b) addition of concentrated amino acid feed solutions, Feed B and Feed C; c) contacting the sample with a Protein A affinity chromatography column; d) selecting at least one wash buffer having pH between 5.8 and 6.2 containing salt and phosphate buffer to minimize aggregation during elution and low pH hold; e) eluting the monoclonal antibody from the Protein A affinity chromatography column with an elution buffer; f) neutralizing protein A eluate to pH 5 by using a neutralization solution having pH between 5.8 and 6.2 devoid of NaOH, instead comprising of citrate buffer in combination with salt; g) subjecting the sample to a second chromatography having cation exchange resin; and wherein the fermentation method maintains low osmolality during logarithmic phase, minimum accumulation of secondary metabolites like ammonia and lactate, provides higher cell growth, high yield followed by a purification method that provides an antibody solution lacking aggregation and devoid of significant turbidity, thereby providing a monomeric antibody with a yield greater than 98% and purity of greater than >99.0 %, having an endotoxin content of <0.10 EU/mg.