Rational design and engineering of therapeutic proteins

Marshall, Shannon; Lazar, Greg A.; Chirino, Arthur J.; Desjarlais, John R.

doi:10.1016/s1359-6446(03)02610-2

Cited by 175 publications

(94 citation statements)

References 61 publications

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“…18,20,21,26,27 In addition to a high specific activity, clinically useful recombinant protein therapeutics must also have suitable protein stability and pharmacokinetic properties. 24,28,29 Toward this end, we here describe singlechain variants of the apoptosis-inducing TRAIL with improved stability and superior product quality in the form of a tumortargeted fusion protein, resulting in high anti-tumor activity in a xenograft tumor model in vivo.…”

Section: Discussionmentioning

confidence: 99%

An optimized antibody-single-chain TRAIL fusion protein for cancer therapy

Siegemund

Seifert

Zarani³

et al. 2016

mAbs

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Fusion proteins combining oligomeric assemblies of a genetically obtained single-chain (sc) variant of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) with antibodies directed against tumorassociated antigens represent a promising strategy to overcome the limited therapeutic activity of conventional soluble TRAIL. To further improve the scTRAIL module in order to obtain a robust, thermostable molecule of high activity, we performed a comprehensive analysis of the minimal TNF homology domain (THD) and optimized linkers between the 3 TRAIL subunits constituting a scTRAIL. Through a stepwise mutagenesis of the N-and C-terminal region and the joining linker sequences, we generated bioactive scTRAIL molecules comprising a covalent linkage of the C-terminal Val280 and the Nterminal position 122 by only 2 amino acid residues in combination with conservative exchanges at positions 122 and 279. The increased thermal stability and solubility of such optimized scTRAIL molecules translated into increased bioactivity in the diabody-scTRAIL (Db-scTRAIL) format, exemplified here for an epidermal growth factor receptor-specific Db-scTRAIL. Additional modifications within the diabody linkers resulted in a fusion protein exerting high, target-dependent apoptosis induction in tumor cell lines in vitro and potent antitumor activity in vivo. Our results illustrate that protein engineering of scTRAIL and associated peptide linkers provides a promising strategy to develop antibody-scTRAIL fusion proteins as effective antitumor therapeutics.

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Section: Discussionmentioning

confidence: 99%

An optimized antibody-single-chain TRAIL fusion protein for cancer therapy

Siegemund

Seifert

Zarani³

et al. 2016

mAbs

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“…Where structural information was incomplete or lacking (e.g., the Fc͞Fc␥RIIb complex), calculations provided a quality set of variants enriched for stability and solubility. The advantage of this capability is because of the innumerable amino acid modifications that are detrimental to proteins (17).…”

Section: Designed Fc Variants Have Optimized Binding Affinity For Fc␥rsmentioning

confidence: 99%

Engineered antibody Fc variants with enhanced effector function

Lazar

Dang²,

Karki³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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702

583

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Antibody-dependent cell-mediated cytotoxicity, a key effector function for the clinical efficacy of monoclonal antibodies, is mediated primarily through a set of closely related Fc␥ receptors with both activating and inhibitory activities. By using computational design algorithms and high-throughput screening, we have engineered a series of Fc variants with optimized Fc␥ receptor affinity and specificity. The designed variants display >2 orders of magnitude enhancement of in vitro effector function, enable efficacy against cells expressing low levels of target antigen, and result in increased cytotoxicity in an in vivo preclinical model. Our engineered Fc regions offer a means for improving the next generation of therapeutic antibodies and have the potential to broaden the diversity of antigens that can be targeted for antibody-based tumor therapy.antibody-dependent cell-mediated cytotoxicity ͉ Fc␥R ͉ protein engineering ͉ cancer

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“…Since initial attempts, the field of protein design has progressed to the point that it is now possible to consider tertiary and quaternary structures and even functional elements for construction of new proteins (28,(37)(38)(39). In this study, we presented the design of engineered GST proteins carrying inserts of the HGK-rich motif derived from a sequence within D5 of HK.…”

Section: Discussionmentioning

confidence: 99%

Chimeric Glutathione S-Transferases Containing Inserts of Kininogen Peptides

Bentley

Merkulov

Peng

et al. 2012

Journal of Biological Chemistry

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Background: Short human kininogen (HK) peptides can be used as therapeutic agents. Results: The biological activity of the HK peptides is dramatically enhanced following insertion into GST. Conclusion: The efficacy of short peptides can be enhanced by insertion into GST without affecting GST structure. Significance: Intra-backbone insertions of small peptides into easily expressed, well characterized soluble proteins may help to create novel protein therapeutics.

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Rational design and engineering of therapeutic proteins

Cited by 175 publications

References 61 publications

An optimized antibody-single-chain TRAIL fusion protein for cancer therapy

An optimized antibody-single-chain TRAIL fusion protein for cancer therapy

Engineered antibody Fc variants with enhanced effector function

Chimeric Glutathione S-Transferases Containing Inserts of Kininogen Peptides

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