Journal ArticleDOI
Construction, bacterial expression and characterization of a bifunctional single-chain antibody-phosphatase fusion protein targeted to the human erbB-2 receptor.
Winfried S. Wels,Ina-Maria Harwerth,Marcus Zwickl,Norman Hardman,Bernd Groner,Nancy E. Hynes +5 more
Reads0
Chats0
TLDR
The bound scFv(FRP5)–PhoA protein can be detected directly on tumor cells using a substrate for alkaline phosphatase, showing that the chimeric protein retains both binding and enzymatic activity.Abstract:
We have constructed genes expressing single-chain antigen binding proteins (scFv) which recognize the human erbB-2 receptor. These genes encode the heavy and light chain variable domains of an erbB-2 receptor specific monoclonal antibody, MAb FRP5, connected by a peptide linker. In order to express a bifunctional molecule, a bacterial alkaline phosphatase gene was fused 3' to the scFv gene. The scFv(FRP5) and scFv(FRP5)-alkaline phosphatase fusion protein (scFv(FRP5)-PhoA) expressed in E. coli specifically recognize the human erbB-2 protein and compete with MAb FRP5 for binding to the receptor. The bound scFv(FRP5)-PhoA protein can be detected directly on tumor cells using a substrate for alkaline phosphatase, showing that the chimeric protein retains both binding and enzymatic activity.read more
Citations
More filters
Patent
Multivalent and multispecific binding proteins, their manufacture and use
Kaspar-Philipp Holliger,Andrew D. Griffiths,Hendricus Renerus Jacobus Matheus Hoogenboom,Magnus Malmqvist,James D. Marks,Brian T. Mcguinness,Anthony Richard Pope,Terence Derek Prospero,Gregory Paul Winter +8 more
TL;DR: In this article, the authors present methods of preparation of polypeptides and multimers and diverse repertoires thereof, and their display on the surface of bacteriophage for easy selection of binders of interest, along with many utilities.
Journal ArticleDOI
Tandem CAR T cells targeting HER2 and IL13Rα2 mitigate tumor antigen escape
Meenakshi Hegde,Malini Mukherjee,Malini Mukherjee,Zakaria Grada,Antonella Pignata,Daniel Landi,Shoba A. Navai,Amanda Wakefield,Kristen Fousek,Kevin Bielamowicz,Kevin Chow,Vita S. Brawley,Tiara T. Byrd,Simone Krebs,Stephen Gottschalk,Winfried S. Wels,Matthew L. Baker,Gianpietro Dotti,Maksim Mamonkin,Malcolm K. Brenner,Jordan S. Orange,Jordan S. Orange,Nabil Ahmed +22 more
TL;DR: TanCAR T cells show therapeutic potential to improve glioblastoma control by coengaging HER2 and IL13Rα2 in an augmented, bivalent immune synapse that enhances T cell functionality and reduces antigen escape.
Journal ArticleDOI
TanCAR: A Novel Bispecific Chimeric Antigen Receptor for Cancer Immunotherapy
Zakaria Grada,Zakaria Grada,Meenakshi Hegde,Meenakshi Hegde,Tiara T. Byrd,Tiara T. Byrd,Donald R. Shaffer,Donald R. Shaffer,Alexia Ghazi,Alexia Ghazi,Vita S. Brawley,Vita S. Brawley,Amanda Corder,Amanda Corder,Kurt Schönfeld,Joachim Koch,Gianpietro Dotti,Gianpietro Dotti,Helen E. Heslop,Stephen Gottschalk,Winfried S. Wels,Matthew L. Baker,Nabil Ahmed,Nabil Ahmed +23 more
TL;DR: TanCAR as mentioned in this paper is a bispecific activation and targeting of T cells that can be used to increase the specificity of effector cells for malignant versus normal target cells, to offset antigen escape or to allow for targeting the tumor and its microenvironment.
METHODS: ORIGINAL ARTICLE TanCAR: A Novel Bispecific Chimeric Antigen Receptor for Cancer Immunotherapy
Zakaria Grada,Meenakshi Hegde,Tiara T. Byrd,Donald R. Shaffer,Alexia Ghazi,Vita S. Brawley,Amanda Corder,Kurt Schönfeld,Joachim Koch,Gianpietro Dotti,Helen E. Heslop,Stephen Gottschalk,Winfried S. Wels,Matthew L. Baker,Nabil Ahmed +14 more
TL;DR: This work created a functional chimeric antigen receptor—the TanCAR, a novel artificial molecule that mediates bispecific activation and targeting of T cells and demonstrates the feasibility of cumulative integration of structure and docking simulation data using computational tools to interrogate the design and predict the functionality of such a complex bispecial molecule.
Journal ArticleDOI
Single-chain antibody-mediated intracellular retention of ErbB-2 impairs Neu differentiation factor and epidermal growth factor signaling.
TL;DR: The observations demonstrate that ErbB-2 plays a central role in the type I/EGFR-related family of receptors and that receptor transmodulation represents a crucial step in growth factor signaling.
References
More filters
Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal ArticleDOI
Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene
Dennis J. Slamon,Gary M. Clark,Steven G. Wong,Wendy J. Levin,Axel Ullrich,William L. McGuire +5 more
TL;DR: Amplification of the HER-2/neu gene was a significant predictor of both overall survival and time to relapse in patients with breast cancer, and had greater prognostic value than most currently used prognostic factors in lymph node-positive disease.
Journal ArticleDOI
Ligand: a versatile computerized approach for characterization of ligand-binding systems.
Peter J. Munson,David Rodbard +1 more
TL;DR: This approach provides two major advantages compared with other available methods: it uses an exact mathematical model of the ligand-binding system, thereby avoiding the possible biases introduced by several commonly used approximations and it uses a statistically valid, appropriately weighted least-squares curve-fitting algorithm with objective measurement of goodness of fit.
Journal ArticleDOI
Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer.
Dennis J. Slamon,William Godolphin,Lovell A. Jones,John A. Holt,Steven G. Wong,Duane E. Keith,Wendy J. Levin,Susan G. Stuart,Judy Udove,Axel Ullrich,Michael F. Press +10 more
TL;DR: The concept that the HER-2/neu gene may be involved in the pathogenesis of some human cancers, including breast and ovarian cancer, is supported.
Journal ArticleDOI
Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli.
James S. Huston,D. Levinson,Meredith Mudgett-Hunter,M S Tai,Jirí Novotný,Michael N. Margolies,R J Ridge,Robert E. Bruccoleri,Edgar Haber,R Crea +9 more
TL;DR: A biosynthetic antibody binding site, which incorporated the variable domains of anti-digoxin monoclonal antibody 26-10 in a single polypeptide chain, was produced in Escherichia coli by protein engineering.
Related Papers (5)
Assembly of a functional immunoglobulin Fv fragment in Escherichia coli.
Arne Skerra,Andreas Plückthun +1 more