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Protein engineering approaches for antibody fragments: directed evolution and rational design approaches - PubMed

  • ️Tue Jan 01 2019

Protein engineering approaches for antibody fragments: directed evolution and rational design approaches

Merve Arslan et al. Turk J Biol. 2019.

Abstract

The number of therapeutic antibodies in preclinical, clinical, or approved phases has been increasing exponentially, mostly due to their known successes. Development of antibody engineering methods has substantially hastened the development of therapeutic antibodies. A variety of protein engineering techniques can be applied to antibodies to improve their afinity and/or biophysical properties such as solubility and stability. Antibody fragments (where all or some parts of constant regions are eliminated while the essential antigen binding region is preserved) are more suitable for protein engineering techniques because there are many in vitro screening technologies available for antibody fragments but not full-length antibodies. Improvement of biophysical characteristics is important in the early development phase because most antibodies fail at the later stage of development and this leads to loss of resources and time. Here, we review directed evolution and rational design methods to improve antibody properties. Recent developments in rational design approaches and antibody display technologies, and especially phage display, which was recently awarded the 2018 Nobel Prize, are discussed to be used in antibody research and development.

Keywords: Antibody; afinity; antibody fragment; biophysical properties; directed evolution; phage display; protein engineering; rational design; yeast surface display.

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Conflict of interest statement

CONFLICT OF INTEREST: none declared

Figures

Figure 1
Figure 1

Antibody display technologies. General schematic for in vitro and in vivo display techniques.

Figure 2
Figure 2

Protein engineering approaches based on antibody regions. While solubility/ aggregation can be improved by engineering exposed residues, stability can be increased by both exposed and core residues. Complementarity-determining regions (CDRs) mainly affect specificity and affinity. Tags/linkers can be added for better functionality.

Figure 3
Figure 3

A case study for web tools. Several tools introduced in this review were used to determine aggregation-prone regions of an scFv sequence used in our lab. Blue highlighted regions are outputs of tools as aggregation prone regions. Each web tool has a different threshold, which was not shown in this figure. Mutation site is selected according to common predicted regions of different tools (at least 6 of 8 tools gave same residues as aggregation-prone).

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