Humanization by variable domain resurfacing and grafting on a human IgG4, using a new approach for determination of non-human like surface accessible framework residues based on homology modelling of variable domains

Staelens, Steven; Desmet, Jan; Ngo, Thu Hoa; Vauterin, Stephan; Pareyn, Inge; Barbeaux, Philippe; Rompaey, Isabel Van; Stassen, Jean–Marie; Deckmyn, Hans; Vanhoorelbeke, Karen

doi:10.1016/j.molimm.2005.07.018

Cited by 35 publications

(26 citation statements)

References 44 publications

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“…The construction of scFv-82D6A3(WT) (European Bioinformatics Institute Database accession number AJ965435) through splicing by overlap extension, its cloning in the pCANTAB 5 E vector, and its expression as a soluble protein in the periplasm of Escherichia coli HB2151 cells has been previously described (52). The scFv-82D6A3(WT) was purified from the periplasmic extract and the supernatant as described, using a HiTrap anti-E Tag column (RPAS purification module; Amersham Biosciences) (52).…”

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

“…The scFv-82D6A3(WT) was purified from the periplasmic extract and the supernatant as described, using a HiTrap anti-E Tag column (RPAS purification module; Amersham Biosciences) (52). After dialysis against PBS, purity was checked in SDS-PAGE using a 10% gel followed by staining with Coomassie Brilliant Blue.…”

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

“…Plasmid DNA was prepared using the NucleoSpin Plasmid Quick Pure kit (Machery-Nagel, Düren, Germany). Sequencing, expression, and purification were performed as described (52). After dialysis against PBS, purity was checked as described above.…”

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

“…Detection of the scFvs with anti-E Tag-horseradish peroxidase antibodies was performed as described (52).…”

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

“…Binding of scFv-82D6A3(WT) and its alanine mutants to VWF was performed as described before (52). Briefly, a 96-well plate was coated with VWF and incubated with dilution series of purified scFv-82D6A3(WT) and its alanine mutants.…”

Section: Binding Of Scfv-82d6a3(wt) and Its Alanine Mutants To Vwf-mentioning

confidence: 99%

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Paratope Determination of the Antithrombotic Antibody 82D6A3 Based on the Crystal Structure of Its Complex with the von Willebrand Factor A3-Domain

Staelens

Hadders

Vauterin

et al. 2006

Journal of Biological Chemistry

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Platelet adhesion, activation, and aggregation are the three major steps in primary hemostasis, a process that is necessary for the cessation of bleeding at sites of vascular injury. The same process is also responsible for pathologic thrombus formation, particularly in arteries at sites of high shear stress. At high shear stress, binding of von Willebrand factor (VWF) 3 to collagen that is exposed in the subendothelial matrix after injury, and the subsequent interaction of the immobilized VWF with the platelet glycoprotein (GP) Ib/IX/V-receptor complex, is a prerequisite for proper adhesion (1-3). As a result of the reversible VWF-GP Ib/IX/V interaction, platelets roll over the injured vessel wall and slow down (3). Subsequent firm adhesion and activation of platelets are mediated by their collagen receptors, integrins ␣2␤1 and GPVI, respectively (4 -8). Activated platelets expose high affinity GPIIb/IIIa receptors that mediate aggregation by binding to fibrinogen and, to a lesser extent, to VWF (9 -11).Collagen type VI and fibrillar collagens types I and III are abundant in the subendothelium (12). VWF interacts with the fibrillar collagens mainly via its A3-domain (13, 14), whereas its A1-domain has been implicated in binding to collagen type VI (15, 16). The latter interaction only mediates platelet adhesion under low shear conditions (17, 18). The A3-domain is homologous in sequence and structure with the I-domains of collagen binding integrin ␣-chains, such as the I-domain of ␣2␤1 (19 -21). These domains adopt a dinucleotide binding fold, or Rossman fold, that consists of a central hydrophobic parallel ␤-sheet flanked on both sites with amphipathic ␣-helices (22-24). Despite their structural similarity, they bind collagen in distinctly different ways. The I-domain of ␣2␤1 binds collagen in a hydrophilic groove at its top face that contains a metal ion-dependent adhesion site (25,26). In contrast, the collagen-binding site of the A3-domain is located at its front face and is rather flat and hydrophobic (23,(27)(28)(29).Our group characterized a monoclonal antibody against human VWF, 82D6A3, that binds with high affinity to the VWF A3-domain and inhibits VWF binding to collagen types I and III (15). Its inhibiting effect on VWF-mediated platelet adhesion becomes more pronounced with increasing shear stress, as is expected because the collagen-VWF-GPIb axis is essential for platelet adhesion under high shear only (30). The discontinuous epitope of 82D6A3, which has recently been unraveled via phage display and binding studies with alanine mutants of the VWF A3-domain, includes several residues of the A3-domain that are important for collagen binding (30). We further demonstrated that 82D6A3 has antithrombotic effects in a baboon arterial thrombosis model (31), proving for the first time that the VWF-collagen interaction is indeed involved in thrombus formation in vivo (32). Interestingly, the antithrombotic effect was not associated with significantly increased bleeding times, even at high doses, demonstrat...

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Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

“…Detection of the scFvs with anti-E Tag-horseradish peroxidase antibodies was performed as described (52).…”

Section: Prokaryotic Expression and Purification Of Wild Type Scfv Ofmentioning

confidence: 99%

Section: Binding Of Scfv-82d6a3(wt) and Its Alanine Mutants To Vwf-mentioning

confidence: 99%

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Paratope Determination of the Antithrombotic Antibody 82D6A3 Based on the Crystal Structure of Its Complex with the von Willebrand Factor A3-Domain

Staelens

Hadders

Vauterin

et al. 2006

Journal of Biological Chemistry

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Therapeutic Monoclonal Antibodies: Past, Present, and Future

Strohl¹

2009

Therapeutic Monoclonal Antibodies

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In this chapter, an overview of the therapeutic antibody industry today, including the many commercial antibodies and Fc fusions and the rich clinical pipeline, is presented and analyzed. The long history of antibodies is given to bring context to the therapeutic antibody industry. This history includes serum therapy, the use of IVIG, and the evolution of those therapies into the development of the monoclonal Therapeutic Monoclonal Antibodies: From Bench to Clinic. Edited by Zhiqiang An Copyright # 2009 John Wiley & Sons, Inc.antibody business as we know it today. The history of technologies that fostered the revolution of therapeutic antibody development in the 1990s is also described. Finally, the future of the therapeutic monoclonal antibody and Fc fusion business is presented along with opportunities and challenges facing the business and those who work in it.Ã Based on published 2005 and 2006 sales, and 2010 projected sales of the top 70 biologics currently on the market, excluding vaccines (multiple sources). ÃÃ Projections were made prior to published safety concerns in mid-2007, which have depressed overall sales of epoetins. INTRODUCTIONÃ Data obtained from prescribing information released by the manufacturers, company websites, Prous Science Integrity. ÃÃConjugate is ozogamicin, a calecheamycin (natural product cytotoxin). ÃÃÃSuspended 2/28/05; reinstated under specified conditions.

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Antibody Engineering: Humanization, Affinity Maturation, and Selection Techniques

Almagro¹,

Strohl²

2009

Therapeutic Monoclonal Antibodies

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Humanization by variable domain resurfacing and grafting on a human IgG4, using a new approach for determination of non-human like surface accessible framework residues based on homology modelling of variable domains

Cited by 35 publications

References 44 publications

Paratope Determination of the Antithrombotic Antibody 82D6A3 Based on the Crystal Structure of Its Complex with the von Willebrand Factor A3-Domain

Paratope Determination of the Antithrombotic Antibody 82D6A3 Based on the Crystal Structure of Its Complex with the von Willebrand Factor A3-Domain

Therapeutic Monoclonal Antibodies: Past, Present, and Future

Antibody Engineering: Humanization, Affinity Maturation, and Selection Techniques

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