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Rapid discovery of lead molecules is critical for accelerating potential biotherapeutics to the clinic but finding those desirable candidates can be challenging. The Opto™ Plasma B Discovery 4.0 workflow enables rapid screening of up to 100,000 B cells to identify 1000s of hits, down-selection of lead molecules by profiling antibody function, and then allows sequencing and re-expression of up to 1,000 functionally characterized antibodies. This key last step in the discovery process — sequence recovery and re-expression — can be costly, time consuming, and laborious. Here our in-house experts discuss some of the major challenges encountered with sequencing and re-expression, and how the Opto Plasma B Discovery 4.0 workflow can help.
Popular methods used for sequencing like Sanger sequencing are cost prohibitive and can only be performed on a limited number of antibodies, while NGS sequencing is less expensive, but is not as accurate for full-length sequences. Once sequences are finally obtained, costly gene synthesis and laborious bacterial cloning can limit the number of lead molecules that are re-expressed.
OptoSeq™ Barcoded BCR offers a less expensive and more efficient method for rapid, accurate sequencing of paired heavy/light chain antibody genes by DNA fragmentation, NGS sequencing, and bioinformatics analysis. This gives you the best of both worlds — the accuracy and full-length sequence information of Sanger at the speed and cost of NGS.
The OptoSeq BCR Rapid Re-expression kit enables re-expression of >1,000 antibodies without gene synthesis and cloning in just one week.
OptoSeq Barcoded BCR enables rapid, accurate sequencing of paired heavy/light chain antibody genes by DNA fragmentation, NGS sequencing, and bioinformatics analysis. Bioinformatic assembly of full-length antibody variable regions from short reads (150 x 150) enables accurate sequencing of thousands of antibodies from a single run on standard Illumina sequencing platforms. Function of re-expressed antibodies can then be confirmed using plate-based ELISA assays.
The OptoSeq BCR Rapid Re-expression kit can convert antibody sequences to molecules in just 1 week for plate-based confirmation assays. Amplified cDNA is cloned into expression constructs in one step, and up to 1,000 antibodies can be expressed for rapid functional confirmation.
Read our Opto Plasma B Discovery 4.0 datasheet to learn more about how OptoSeq Barcoded BCR and OptoSeq BCR Rapid Re-expression kits can be used to sequence and re-express up to 1,000 functionally characterized antibodies in less than 1 week.
This story originally appeared in the Spring/Summer 2021 edition of BLI News. To read more, check out our latest edition.