Single domain antibodies (sdAbs or VHHs) offer the affinity and therapeutic value of conventional antibodies with increased stability and solubility. Unlike conventional antibodies, however, VHHs do not benefit from a platform production process. Successful VHH production has been shown in numerous microbial hosts including Escherichia coli and Pichia pastoris. Typical purification processes, however, either rely on affinity tags or are unique for each new product. Affinity tags are often a cause of concern for regulators, while developing a brand new process for each product requires significant time and resources. A ‘plug and play’ process for the production and purification of VHHs could simplify the manufacturing of these products and enable the rapid translation from sequence to pre-clinical material. We have previously demonstrated a bench-scale, integrated and automated system capable of rapidly producing a variety of high quality biologic drugs. Here we use this system to demonstrate a broadly applicable production and purification process for single domain antibodies in Pichia pastoris which enables the rapid production of multiple VHH products at a quality appropriate for pre-clinical studies. This process includes fermentation via perfusion, a single in-line pH adjustment, and a two-step, integrated chromatographic purification. To date, we have successfully produced more than five different VHH products using this production process with only minimal process adjustments. Further, we show that the necessary process adjustments can be predicted from the target molecule sequence, often with no experimental evaluation. We use this methodology to produce pre-clinical quality VHH products in as few as 4 weeks after obtaining the target product sequence. Rapid translation from sequence to pre-clinical quality material as presented could allow a greater number of higher quality molecules to reach the clinic.

Development of a platform process for the rapid production and purification of single domain antibodies

2019 Spring National Meeting

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