The scaling-up process was consistent with the results of the optimized process, further demonstrating the feasibility of this method. The anti-NGF Nb that were further purified using Capto S ImpAct achieved 84.5% yield and 99.2% purity and 77 ppm of HCP. The anti-NGF Nb captured by Capto MMC was 67.2% yield, 94.5% monomer purity, and host cell protein (HCP) was reduced from 74,931 ppm to 484 ppm. Additionally, direct capture of anti-NGF Nb using Capto MMC without adjusting anti-NGF Nb harvest pH was investigated. Furthermore, high purity and yield products can be obtained at laboratory scale and commercial production scale by adjusting the protein pH.
First, pH 4.0-5.0 was demonstrated to be the optimal loading condition for Capto MMC to capture anti-NGF by the design of experiments (DOE). Another possible reason for the change in morphology could be unforeseen effects of the critical point drying process on aged Capto Adhere resin. In this study, a method for capture and purification of an untagged Nb (anti-NGF Nb) by mixed weak cation chromatography and cation exchange chromatography was established. Fig 1.Sketch showing the strong ion exchange group of Capto S ImpAct The medium is designed for intermediate purification or polishing. Nerve growth factor (NGF), which regulates the growth and development of peripheral and central neurons, maintains neuronal survival and plays a key role in both arthritis and acute and chronic pain. 2 Properties of Capto S ImpAct The structure of the well established S (sulfonate group) ligand used for Capto S ImpAct is shown in Figure below. Typically, Nbs are captured by affinity chromatography via the addition of specific fusion tags at their N or C terminus. Nanobodies (Nbs) are single-domain antibodies, which have potential application value in tumor-targeted therapy, immunotherapy, diagnostic probe, and molecular imaging.
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