Stepwisein vitroaffinity maturation of Vitaxin, an αvβ3-specific humanized mAb

Wu, Herren; Beuerlein, Gregory; Nie, Ying; Smith, H. S.; Lee, Bruce A.; Hensler, Mary E.; Huse, William D.; Watkins, Jeffry D.

doi:10.1073/pnas.95.11.6037

Cited by 138 publications

(110 citation statements)

References 33 publications

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“…Although these strategies can result in significant affinity enhancement, none comprehensively interrogate all six CDR positions in a precise and informative fashion. Other stochastic methods used for maturation (21,23) are also limited because of the inherent randomness in such mutagenesis strategies. Although improved properties might be obtained through these methods, a directed, rational, and informative method based on chemical principles should yield vastly improved variants and informative results about the binding site architecture.…”

Section: Discussionmentioning

confidence: 99%

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A general method for greatly improving the affinity of antibodies by using combinatorial libraries

Rajpal

Beyaz

Haber

et al. 2005

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

144

102

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Look-through mutagenesis (LTM) is a multidimensional mutagenesis method that simultaneously assesses and optimizes combinatorial mutations of selected amino acids. The process focuses on a precise distribution within one or more complementarity determining region (CDR) domains and explores the synergistic contribution of amino acid side-chain chemistry. LTM was applied to an anti-TNF-␣ antibody, D2E7, which is a challenging test case, because D2E7 was highly optimized (Kd ‫؍‬ 1 nM) by others. We selected and incorporated nine amino acids, representative of the major chemical functionalities, individually at every position in each CDR and across all six CDRs (57 aa). Synthetic oligonucleotides, each introducing one amino acid mutation throughout the six CDRs, were pooled to generate segregated libraries containing single mutations in one, two, and͞or three CDRs for each V H and VL domain. Corresponding antibody libraries were displayed on the cell surface of yeast. After positive binding selection, 38 substitutions in 21 CDR positions were identified that resulted in higher affinity binding to TNF-␣. These beneficial mutations in both VH and VL were represented in two combinatorial beneficial mutagenesis libraries and selected by FACS to produce a convergence of variants that exhibit between 500-and 870-fold higher affinities. Importantly, these enhanced affinities translate to a 15-to 30-fold improvement in in vitro TNF-␣ neutralization in an L929 bioassay. Thus, this LTM͞combinatorial beneficial mutagenesis strategy generates a comprehensive energetic map of the antibody-binding site in a facile and rapid manner and should be broadly applicable to the affinity maturation of antibodies and other proteins.look-through mutagenesis ͉ maturation ͉ mutagenesis ͉ TNF-␣

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Section: Discussionmentioning

confidence: 99%

“…Numerous protein engineering and mutagenesis strategies have been used for the enhancement of antibody affinity (3,5,6,11,(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29). For small regions (Ͻ8-10 aa), saturation mutagenesis is typically used to produce all combinations of the 20 naturally occurring amino acids.…”

Section: Discussionmentioning

confidence: 99%

A general method for greatly improving the affinity of antibodies by using combinatorial libraries

Rajpal

Beyaz

Haber

et al. 2005

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

144

102

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“…In the subsequent Phase I clinical trial [Gutheil et al, 2000], Vitaxin I has been proven safe for therapeutic use, however, it had limited efficacy in patients with advanced malignancies. The affinity of this antibody was further increased using stepwise maturation with phage expression libraries [Wu et al, 1998]. The resulting antibody was named Abegrin (Vitaxin II, MEDI-522), which was later licensed to MedImmune, Inc. (Gaithersburg, MD).…”

Section: Integrin α V β 3 Inhibitorsmentioning

confidence: 99%

Integrin α_vβ₃‐targeted cancer therapy

Liu

Wang

Chen

2008

Drug Development Research

281

217

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“…[6][7][8][11][12][13][14][15] Etaracizumab (Abegrin or MEDI-522; MedImmune, Gaithersburg, Md) is an IgG1 humanized monoclonal antibody engineered from the murine monoclonal LM609, an antibody directed against a conformational epitope of alpha v beta 3 (a v b 3 ) integrin that is present on the surface of certain types of invasive tumor cells, angiogenic endothelial cells, and mature osteoclasts. 16 Although it is expressed at low levels in most normal tissues, a v b 3 integrin is highly expressed in certain malignancies, such as melanoma, late-stage glioma, and renal cell carcinoma, as well as a limited number of cell types, such as endothelial cells involved in angiogenesis and mature osteoclasts involved in bone resorption. [17][18][19][20][21] Preclinical studies using a v b 3 antagonists have reported inhibition of melanoma tumor growth independent of its antiangiogenic effects.…”

mentioning

confidence: 99%

A randomized phase 2 study of etaracizumab, a monoclonal antibody against integrin α_vβ₃, ± dacarbazine in patients with stage IV metastatic melanoma

Hersey¹,

Sosman²,

O’Day³

et al. 2010

Cancer

173

115

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BACKGROUND:The alpha v beta 3 (a v b 3 ) integrin is involved in intracellular signaling regulating cell proliferation, migration, and differentiation and is important for tumor-induced angiogenesis. METHODS: This phase 2, randomized, open-label, 2-arm study was designed to capture safety data and evaluate the antitumor efficacy of etaracizumab (Abegrin), an IgG1 humanized monoclonal antibody against the a v b 3 integrin, in patients with previously untreated metastatic melanoma. The objective was to evaluate whether etaracizumab AE dacarbazine had sufficient clinical activity to warrant further study in a phase 3 clinical trial. RESULTS: One hundred twelve patients were randomized to receive etaracizumab alone (N ¼ 57) or etaracizumab þ dacarbazine (N ¼ 55). Safety of etaracizumab AE dacarbazine was acceptable with infusion-related, gastrointestinal, and metabolic reactions being the most common adverse events (AEs). The majority of AEs were grade 1 or 2 in severity in both study arms; most events were not considered serious, except for cardiovascular (myocardial infarction, atrial fibrillation) and thromboembolic events, which occurred in 3 and 5 patients, respectively. None of the patients in the etaracizumab-alone study arm and 12.7% of patients in the etaracizumab þ dacarbazine study arm achieved an objective response. The median duration of objective response in the etaracizumab þ dacarbazine study arm was 4.2 months. Stable disease rate, time to progression (TTP), and progression-free survival (PFS) appeared to be similar between the 2 treatment arms. Stable disease occurred in 45.6% of patients in the etaracizumab-alone study arm and 40.0% of patients in the etaracizumab þ dacarbazine study arm. Median TTP and median PFS were both 1.8 months in the etaracizumab-alone study arm and 2.5 and 2.6 months in the etaracizumab þ dacarbazine study arm, respectively. Median overall survival was 12.6 months in the etaracizumab-alone study arm and 9.4 months in the etaracizumab þ dacarbazine study arm. CON-CLUSIONS: The survival results in both treatment arms of this study were considered unlikely to result in clinically meaningful improvement over dacarbazine alone. Cancer 2010;116:1526-34.

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Stepwisein vitroaffinity maturation of Vitaxin, an α_vβ₃-specific humanized mAb

Cited by 138 publications

References 33 publications

A general method for greatly improving the affinity of antibodies by using combinatorial libraries

A general method for greatly improving the affinity of antibodies by using combinatorial libraries

Integrin α_vβ₃‐targeted cancer therapy

A randomized phase 2 study of etaracizumab, a monoclonal antibody against integrin α_vβ₃, ± dacarbazine in patients with stage IV metastatic melanoma

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Stepwisein vitroaffinity maturation of Vitaxin, an αvβ3-specific humanized mAb

Cited by 138 publications

References 33 publications

A general method for greatly improving the affinity of antibodies by using combinatorial libraries

A general method for greatly improving the affinity of antibodies by using combinatorial libraries

Integrin αvβ3‐targeted cancer therapy

A randomized phase 2 study of etaracizumab, a monoclonal antibody against integrin αvβ3, ± dacarbazine in patients with stage IV metastatic melanoma

Contact Info

Product

Resources

About

Stepwisein vitroaffinity maturation of Vitaxin, an α_vβ₃-specific humanized mAb

Integrin α_vβ₃‐targeted cancer therapy

A randomized phase 2 study of etaracizumab, a monoclonal antibody against integrin α_vβ₃, ± dacarbazine in patients with stage IV metastatic melanoma