3in1: an ultra-fast phage display biopanning method overcoming the biopanning bias and yielding more diverse binders.

Phage display combined with biopanning is a powerful method for the discovery of therapeutic antibodies. Classic biopanning is lengthy and labor-intensive, requiring at least five days of work. The biopanning process is plagued by a bias toward binders with propagation advantages, causing a loss in diversity. We present a more efficient biopanning technique named 3in1 biopanning. Using non-denaturing elution and skipping the intermediary phage amplification steps, 3in1 biopanning performs all biopanning rounds in a single day and overcomes the inherent biopanning bias. 3in1 is five times faster than conventional biopanning and yields up to six times more unique hits with a wide range of affinities. It shortens lead discovery time by a week and reduces consumables and waste. We demonstrate that this novel biopanning technique is suitable for naive-synthetic and immune libraries against a variety of targets.