2017
DOI: 10.1038/srep45259
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Efficient affinity maturation of antibody variable domains requires co-selection of compensatory mutations to maintain thermodynamic stability

Abstract: The ability of antibodies to accumulate affinity-enhancing mutations in their complementarity-determining regions (CDRs) without compromising thermodynamic stability is critical to their natural function. However, it is unclear if affinity mutations in the hypervariable CDRs generally impact antibody stability and to what extent additional compensatory mutations are required to maintain stability during affinity maturation. Here we have experimentally and computationally evaluated the functional contributions … Show more

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Cited by 85 publications
(89 citation statements)
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References 80 publications
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“…Thus, polar sites separated by ∼ 1 nm can pin water effectively in the region between them, and, similarly, a polar or charged site has a larger impact on hydrophobicity and interactions when placed at the center of a hydrophobic patch instead of at its periphery. Such context dependence will play an important role in protein engineering [for example, in engineering of antibodies to optimize both affinity and specificity (66)] as well as in materials design.…”
Section: Discussionmentioning
confidence: 99%
“…Thus, polar sites separated by ∼ 1 nm can pin water effectively in the region between them, and, similarly, a polar or charged site has a larger impact on hydrophobicity and interactions when placed at the center of a hydrophobic patch instead of at its periphery. Such context dependence will play an important role in protein engineering [for example, in engineering of antibodies to optimize both affinity and specificity (66)] as well as in materials design.…”
Section: Discussionmentioning
confidence: 99%
“…The importance of affinity/stability trade-offs during antibody affinity maturation was recently highlighted for single-domain (V H ) antibodies [13, 14]. The investigators introduced mutations throughout the V H frameworks and CDRs using error-prone PCR and displayed the antibody libraries on the surface of yeast.…”
Section: Antibody Affinity/stability Trade-offsmentioning
confidence: 99%
“…Therefore, it will be especially important to generate guidelines for predicting antibody CDR sequences that maximize several biophysical properties (e.g., specificity and solubility) without eliminating key interactive residues that contribute to antibody affinity. The library design methods should also consider amino acid diversity in antibody frameworks (in addition to the CDRs) to enable the identification of both affinity-enhancing mutations (that can be destabilizing) and compensatory stabilizing mutations [13, 14]. Previous work suggests that library design methods that follow patterns of natural antibody diversity for both framework regions [71] and CDRs [31, 7274] may be most effective.…”
Section: Future Directionsmentioning
confidence: 99%
“…These antibodies have undergone B-cell selection for target binding and stability [4, 5] and later for eliciting an in vitro and in vivo biological effect capable of altering a disease pathway. The direct use of the resulting antibodies is not possible due to immune reactions and subsequent formation of antidrug antibodies (ADAs) upon administration into patients.…”
Section: Introductionmentioning
confidence: 99%
“…It is important, however, to consider other biophysical properties that impact the ability to develop an antibody as a therapeutic such as thermal stability [4], nonspecific binding and clearance [23], and propensity to aggregate in solution [24], as well as solubility, FcRn binding, and dissociation. Antibody mutations arising from in vitro somatic hypermutation, as with in vivo B-cell maturation, fall within CDRs, in particular heavy chain CDR1 and CDR2 at the antigen-contacting residues highlighted in Fig.…”
mentioning
confidence: 99%