Abstract
The poor biophysical properties of antibody fragments such as scFvs and diabodies can preclude their use as therapeutic agents. The non-ideal biophysical properties and insufficient thermal stability of antibody fragments often leads to poor expression, poor solubility, and a predisposition of the proteins to aggregate. We have developed a general platform for engineering stability into antibody fragments. By promoting Escherichia coli cultures to secrete scFvs directly into growth media, automated screening methods can be applied to empirically evaluate multiple stability design strategies including rational, sequence-based, and structure-based designs. Once stabilized, these antibody fragments demonstrate improved expression and durability during purification, handling, and storage. Stabilized antibody fragments can also be used as building blocks for multivalent or bispecific antibody-like molecules.
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Acknowledgments
The authors would like to thank Ina Hughes, Xuifeng Wu, and Flora Huang for technical assistance with these experiments.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Miller, B.R., Glaser, S.M., Demarest, S.J. (2009). Rapid Screening Platform for Stabilization of scFvs in Escherichia coli . In: Dimitrov, A. (eds) Therapeutic Antibodies. Methods in Molecular Biology™, vol 525. Humana Press. https://doi.org/10.1007/978-1-59745-554-1_14
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DOI: https://doi.org/10.1007/978-1-59745-554-1_14
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