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Now ready - Easy Bispecific Antibody Evaluation

Analyze specific binding of bsAbs in addition to IgG titer in a single high-throughput assay

Eva Rubio-Marrero, Danyang Huang and Girish Pendse, Celgene Corporation, Summit, NJ Hongshan Li and Sriram Kumaraswamy, FortéBio, Fremont, CA

The ease of use and rapid processing of samples makes the Octet® HTX system an excellent option for higher throughput applications such as ex-pression clone screening and bispecific molecule evaluation. By combining the power of Dip and Read™ biosensors with the Octet® HTX system, one can minimize time, effort, and costs in the assay development of bispecific therapeutics.

Example workflow for bsAb quantitative assay. The assay consists of five assay steps. Step 1: equilibration, Step 2: loading and capture of His-tagged antigen, Step 3: baseline, Step 4. bsAb association, Step 5: bsAb dissociation.

Example of complete Octet® assay run for the association of bsAb to a specific antigen. Lower yellow curve represents negative control where anti-gen was not loaded to the biosensor and 100 µg/mL of bsAb tested for binding.

The production of bispecific antibodies adds a challenge for scientists in the cell line screening process since many other conformations of antibodies can be produced by the cells. If screening is based only on Protein A titer, good pools and clone candidates might be discarded. For this reason, a high-throughput assay using the Octet® HTX system to identify pools and clones with higher concentrations of bispecific antibodies was developed.

This novel BLI approach offers an easy screening method and workflow that assesses bsAb interactions in a versatile, label-free, and easy-to-use format.

There is currently a limitation in technologies that allow for quantitative functional assessment of two interactions to one bispecific molecule in the market. The AppNote shows an easy method for bsAb evaluation. Utilizing Protein A and HIS1K biosensors to identify conditions quickly that are suitable for quantitative measurement of bispecific binding.

By analyzing specific binding results in addition to IgG titer in a single high-throughput instru-ment, the binding activity of a bispecific molecule to both targets can be assessed to rank the top pools/clones during cell line development. The results obtained from the Octet binding assay shown have been validated using other analytical methods such as RP-HPLC, CE-SDS, SPR, and LC-MS to show that binding results can be correlated to the purity of the bsAb from the samples tested.

Facts, background information, dossiers
  • cell line screening
  • bispecific antibodies
  • protein-protein interaction
  • biolayer interferometry
  • Protein A
  • high-throughput assays
  • antibody screening
  • IgG titer
  • concentration measurement
More about FortéBio
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