Fab-based bispecific antibody formats with robust biophysical properties and biological activity
| Autor: | Arlene Sereno, Brian Kuhlman, Xiufeng Wu, F. Huang, Micheal A. Batt, Steven M. Lewis, Andrew L. Glasebrook, Ricky Lieu, Cody Fine, Carina Torres, S.J. Demarest, J.R. Fitchett, Caroline Weldon |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Bispecific antibody
Protein Stability Chemistry Stereochemistry Recombinant Fusion Proteins Immunology Biological activity Computational biology Protein engineering Immunoglobulin light chain Protein stability Cell Line Tumor Antibodies Bispecific Humans Immunology and Allergy Single-Chain Antibodies Reports |
| DOI: | 10.17615/3vv5-xm53 |
| Popis: | A myriad of innovative bispecific antibody (BsAb) platforms have been reported. Most require significant protein engineering to be viable from a development and manufacturing perspective. Single-chain variable fragments (scFvs) and diabodies that consist only of antibody variable domains have been used as building blocks for making BsAbs for decades. The drawback with Fv-only moieties is that they lack the native-like interactions with CH1/CL domains that make antibody Fab regions stable and soluble. Here, we utilize a redesigned Fab interface to explore 2 novel Fab-based BsAbs platforms. The redesigned Fab interface designs limit heavy and light chain mixing when 2 Fabs are co-expressed simultaneously, thus allowing the use of 2 different Fabs within a BsAb construct without the requirement of one or more scFvs. We describe the stability and activity of a HER2×HER2 IgG-Fab BsAb, and compare its biophysical and activity properties with those of an IgG-scFv that utilizes the variable domains of the same parental antibodies. We also generated an EGFR × CD3 tandem Fab protein with a similar format to a tandem scFv (otherwise known as a bispecific T cell engager or BiTE). We show that the Fab-based BsAbs have superior biophysical properties compared to the scFv-based BsAbs. Additionally, the Fab-based BsAbs do not simply recapitulate the activity of their scFv counterparts, but are shown to possess unique biological activity. |
| Databáze: | OpenAIRE |
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