A fully human antibody neutralising biologically active human TGFβ2 for use in therapy

Thompson, Julia; Vaughan, Tristan J.; Williams, Andrew; Wilton, Jane; Johnson, Kevin S.; Bacon, Louise; Green, Jonathan A.; Field, Ray; Ruddock, Steven; Martins, Mia; Pope, Anthony R.; Tempest, Philip R.; Jackson, Ronald H.

doi:10.1016/s0022-1759(99)00060-5

Cited by 45 publications

(16 citation statements)

References 62 publications

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“…The purified PSG2 scFv clone was identified as an scFv fragment whose binding to 19.ek.Fc was inhibited in the presence of increasing concentrations of the murine PSGL-1/Fc fusion protein, which contains sulfated tyrosines within a different sequence context (34). The PSG2 scFv fragment was converted to a full-length intact IgG 4 antibody (designated PSG2) and expressed in mammalian Chinese hamster ovary cells as described (35). PSG2 Purification-PSG2-expressing Chinese hamster ovary cells were expanded from a single vial of frozen cells in ␣-minimal essential medium (Mediatech, Inc.) containing 10% heatinactivated dialyzed fetal bovine serum (Sigma), 2 mM L-glutamine, 100 nM methotrexate, 1 mg/ml G418, 100 units/ml penicillin, and 100 g/ml streptomycin into 850-cm 2 roller bottles.…”

Section: Methodsmentioning

confidence: 99%

Detection and Purification of Tyrosine-sulfated Proteins Using a Novel Anti-sulfotyrosine Monoclonal Antibody

Hoffhines¹,

Damoc

Bridges³

et al. 2006

Journal of Biological Chemistry

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Protein tyrosine O-sulfation is a post-translational modification mediated by one of two Golgi tyrosylprotein sulfotransferases (TPST1 and TPST2) that catalyze the transfer of sulfate to tyrosine residues in secreted and transmembrane proteins. Tyrosine sulfation plays a role in protein-protein interactions in several well defined systems. Although dozens of tyrosine-sulfated proteins are known, many more are likely to exist and await description. Advancing our understanding of the importance of tyrosine sulfation in biological systems requires the development of new tools for the detection and study of tyrosine-sulfated proteins. We have developed a novel anti-sulfotyrosine monoclonal antibody (called PSG2) that binds with high affinity and exquisite specificity to sulfotyrosine residues in peptides and proteins independently of sequence context. We show that it can detect tyrosine-sulfated proteins in complex biological samples and can be used as a probe to assess the role of tyrosine sulfation in protein function. We also demonstrate the utility of PSG2 in the purification of tyrosine-sulfated proteins from crude tissue samples. Finally, Western blot analysis using PSG2 showed that certain sperm/epididymal proteins are undersulfated in Tpst2 ؊/؊ mice. This indicates that TPST1 and TPST2 have distinct macromolecular substrate specificities and provides clues as to the molecular mechanism of the infertility of Tpst2 ؊/؊ males. PSG2 should be widely applicable for identification of tyrosine-sulfated proteins in other systems and organisms.Protein tyrosine O-sulfation is a post-translational modification that occurs in most eukaryotes (1-3). In mouse and man, tyrosine sulfation is mediated by one of only two tyrosylprotein sulfotransferases (EC 2.8.2.20), TPST1 and TPST2 (4 -6). These enzymes catalyze the transfer of sulfate from 3Ј-phosphoadenosine 5Ј-phosphosulfate, the universal sulfate donor, to tyrosine residues in polypeptides (7). TPST enzymes are type II transmembrane proteins that reside in the trans-Golgi network and have luminally oriented catalytic domains (4 -6, 8). Thus, tyrosine sulfation occurs only on soluble and transmembrane proteins that transit the Golgi en route to either secretion or incorporation into the plasma membrane. Accordingly, all of the native tyrosine-sulfated proteins described to date fall into one of these two categories (2).Consensus features for tyrosine sulfation have been proposed based on the amino acid sequences flanking known sulfation sites coupled with in vitro studies on the sulfation of various synthetic peptides (PROSITE accession number PS00003). In addition, a software tool for prediction of tyrosine sulfation sites in proteins called Sulfinator has been developed (9). However, the positive predictive value of these features and the Sulfinator tool is not known. Some known tyrosine sulfation sites do not fulfill proposed consensus features, and some are not predicted by Sulfinator. Thus, unlike some other posttranslational modifications, there is no way to reliab...

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Section: Methodsmentioning

confidence: 99%

Detection and Purification of Tyrosine-sulfated Proteins Using a Novel Anti-sulfotyrosine Monoclonal Antibody

Hoffhines¹,

Damoc

Bridges³

et al. 2006

Journal of Biological Chemistry

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“…Data from Biswas et al, presented in abstract form, also showed that 1D11 reduced skeletal tumor burden and osteolytic bone lesions caused by MDA-MB-231 cells in mice while also increasing the bone volume [117]. For use in patients, fully humanized TGF-β monoclonal neutralizing antibodies have been developed, including Lerdelimumab/CAT-152 [118,119], Metelimumab/CAT-192 [120] and GC-1008 [9]. The TGF-β 2 neutralizing antibody Lerdelimumab was developed by Cambridge Antibody Technology (CAT) and has been successfully used in phase I/II clinical trials to reduce scarring after glaucoma surgery [121].…”

Section: Tgf-β As Therapeutic Targetmentioning

confidence: 99%

TGF-β in the Bone Microenvironment: Role in Breast Cancer Metastases

Buijs

Stayrook

Guise

2011

Cancer Microenvironment

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Breast cancer is the most prevalent cancer among females worldwide. It has long been known that cancers preferentially metastasize to particular organs, and bone metastases occur in ∼70% of patients with advanced breast cancer. Breast cancer bone metastases are predominantly osteolytic and accompanied by bone destruction, bone fractures, pain, and hypercalcemia, causing severe morbidity and hospitalization. In the bone matrix, transforming growth factor-β (TGF-β) is one of the most abundant growth factors, which is released in active form upon tumor-induced osteoclastic bone resorption. TGF-β, in turn, stimulates bone metastatic cells to secrete factors that further drive osteolytic destruction of the bone adjacent to the tumor, categorizing TGF-β as a crucial factor responsible for driving the feed-forward vicious cycle of cancer growth in bone. Moreover, TGF-β activates epithelial-to-mesenchymal transition, increases tumor cell invasiveness and angiogenesis and induces immunosuppression. Blocking the TGF-β signaling pathway to interrupt this vicious cycle between breast cancer and bone offers a promising target for therapeutic intervention to decrease skeletal metastasis. This review will describe the role of TGF-β in breast cancer and bone metastasis, and pre-clinical and clinical data will be evaluated for the potential use of TGF-β inhibitors in clinical practice to treat breast cancer bone metastases.

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“…Given its key function in fibrosis development, TGF-β signaling has been a primary therapeutic target that can be modulated directly with antibodies such as lerdelimumab, soluble TGF-β1 receptors, or small molecules that attenuate TGF-β downstream signaling (120)(121)(122)(123)(124). However, systemic TGF-β inhibition increased inflammatory responses, so simple blockade of these key elements might not be effective because of the complexity of fibrogenesis (119,125).…”

Section: Similar Pathophysiological Concepts Of Nash and Ash With CLImentioning

confidence: 99%

Therapeutic opportunities for alcoholic steatohepatitis and nonalcoholic steatohepatitis: exploiting similarities and differences in pathogenesis

et al. 2017

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A fully human antibody neutralising biologically active human TGFβ2 for use in therapy

Cited by 45 publications

References 62 publications

Detection and Purification of Tyrosine-sulfated Proteins Using a Novel Anti-sulfotyrosine Monoclonal Antibody

Detection and Purification of Tyrosine-sulfated Proteins Using a Novel Anti-sulfotyrosine Monoclonal Antibody

TGF-β in the Bone Microenvironment: Role in Breast Cancer Metastases

Therapeutic opportunities for alcoholic steatohepatitis and nonalcoholic steatohepatitis: exploiting similarities and differences in pathogenesis

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