Resources Webinars Biointron x AET US 2023 Webinar: Accelerating Early Discovery through HTP & High-Speed Antibody Production

Biointron x AET US 2023 Webinar: Accelerating Early Discovery through HTP & High-Speed Antibody Production

Biointron 2024-03-21 Read time: 10 mins

Biointron's SVP of R&D, Lei Shi, PhD, discussed the processes of accelerating early discovery through HTP and high-speed antibody production during the Antibody Engineering & Therapeutics US 2023 conference in San Diego. Below is a summary of the talk.

Focused on Antibody Discovery: From Lead Generation to Cell Line Development

Biointron has the capability of high-throughput antibody production in only 1-2 weeks for any isotype and any species. Our process innovation and automation streamlined production involves cell dispensing, plasmid preparation, and pipetting and labeling. This year, we made >50,000 monoclonal antibodies (mAbs) in various scales, with robust expression and high purity of mAb(-like) production. 

Case 1: 818 IgG production in a single batch 

  • Client engaged Biointron to express a panel of MAbs with HTP service 

  • Text-to-Test: The production of all 818 mAbs were delivered in 10 days in 1 single batch 

  • Saved time to focus on downstream assay 

Case 2: Continuing improvement of VHH-Fc production 

  • Average final yield is 2.18mg 180 Vhh-Fc, single batch, 10ml 

  • 24-well HTP expression in 4 days → provides faster sample production 

Extensive Experiences in Bispecific (BsAb) Formats

>40,000 bsAbs are made annually, and they are custom-produced with multiple formats available. Purified bsAb products can be developed in as fast as 2-3 weeks. KIH (knobs-into-holes) formats can also be expressed with either 1-step or 2-step purification. 

Other Variants

  • IgG-Vhh: Antibodies can be symmetrical or asymmetrical. 

  • IgG-scFv: Antibodies can be symmetrical or extended. 

FC-MES™ Affinity Maturation: The 6-Week Workflow

This HTP process starts with no-bias 17 codon primers mixture to generate 17 amino acid mutations. After generating all 17 aa variants, we transform 17 amino acid mutation mixture and pick up 88 colonies. HTP expression then occurs in 96 well plate and binding is measured by ELISA with supernatants. We can also combine the mutations to improve affinity. 

Alpaca VHH Discovery Platform

Biointron owns an alpaca farm with >350 animals. We utilize an immunization strategy to isolate PBMCs for either phage display (1.5 month/high-throughput), single B cell sorting (3-4 weeks), or NGS sequencing (2-3 weeks). 

VHH Discovery Case for Anti-X Nanobody 

  • Target X, a cell surface receptor expressed on T cells, is a co-stimulator for T cell activation. 

  • From the alpaca immune antibody library, phage display tech was used to screen the nanobodies targeting X. 

  • The selected hits were functionally characterized & validated. 

  • The candidate sequences have been selected. Humanization & affinity maturation are being carried out in preparation for the final construction of the bispecific antibody. 

  • Result: 49 VHH Antibodies Bound to X in SPR; Affinity range 5.36x10-7 M to 7.66x10-10 M. 

Single B Cell Sorting Discovery Platform

A microfluidics platform is used to sort oil-based droplets, and then combined with NGS sequencing and HTP expression to identify hits in 1 week, and we get binder data in 3-4 weeks after humanization.

To learn more, please contact Biointron’s expert team at info@biointron.com, or call +1 (732) 790-8340.

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