neoSwitch™ enables binder discovery and protein production in the same host under control of a chemical trigger in the growth media, eliminating weeks of subcloning and protein expression in a secondary host which is normally required to obtain soluble protein for characterization (SPR, ELISA etc). neoSwitch comes pre-transformed with high diversity libraries for ‘plug and play’ yeast display workflows.

Figure 1: neoSwitch ‘switches’ between display and secretion with a media change

Validation of neoSwitch

After transforming neoSwitch™ with Neo’s synthetic Llama VHH library, 12 clones were randomly selected to measure surface display by flow cytometry with an anti-Myc secondary. The majority of clones expressed at levels comparable to a validated control VHH. Next, the clones were grown in 96-well plates for 44 hours at 25˚C in specialty media. 6/12 clones secreted detectable protein as measured by PAGE gel of raw supernatant. These data demonstrate the potential of neoSwitch™ to accelerate antibody discovery workflows and identify high performing clones in a few days instead of the weeks required to do the same in standard workflows.

Figure 3: Stain free PAGE-gel of supernatant. Ten randomly selected clones (green) were compared to positive (black, left) and negative (black, right) controls. 6 clones had measurable secretion and one clone was equivalent to the positive control (~20mg/L).

Iterative campaigns to generate diversity from top hits

% of population display
70% (comparable to EBY100)
Pre-transformed
Aliquots of yeast transformed with Neo’s synthetic naive VHH library or customer library at high diversity (eg. 109)
Protein Yield
Validated 300nM average VHH yield from random library
Protein Production Timeline
Obtain protein for downstream assays in ~72 hours

Product Features

Pre-transformed Library
Neo’s synthetic VHH library
Description
Synthetic naive library with Llama framework
Time to Delivery
Ships immediately

Library Specifications and TAT

Neo Designed Synthetic VHH Library Llama VHH Library Ships Immediately
Customer-Defined Library Custom Mutational Schema Subject to Library Build Time
Customer Existing Starting Material e.g. cDNA from immunization, pre-existing library Subject to Library Build Time

Synthetic Naive Library for Antibody Discovery

Library Size
1.3 x 109
Backbone
Llama framework to enable high affinity binder discovery
CDR Diversity
  • Controlled mutagenesis library biased for CDRs to create functional binders
  • CDR3 is a mixture of 3 different lengths
  • Sequence liabilities (e.g. cysteine and methionine) are removed
Library Validation
Library diversity validated in neoDisplay by NGS; amino acid frequency matches design
Library Performance Validation
We performed a discovery campaign against human PD1 and discovered 33 unique binders (defined as CDR3 with at least 2 AA differences from any other sequence in set)

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