| The use of phage display to generate conformation-sensor recombinant antibodies. | |
| | |
MedLine Citation:
|
PMID: 23154784 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
We describe a phage display approach that we have previously used to generate conformation-sensor antibodies that specifically recognize and stabilize the oxidized, inactive conformation of protein tyrosine phosphatase 1B (PTP1B). We use a solution-based panning and screening strategy conducted in the presence of reduced active PTP1B, which enriches antibodies to epitopes unique to the oxidized form while excluding antibodies that recognize epitopes common to oxidized and reduced forms of PTP1B. This strategy avoids conventional solid-phase immobilization owing to its inherent potential for denaturation of the antigen. In addition, a functional screening strategy selects single-chain variable fragments (scFvs) directly for their capacity for both specific binding and stabilization of the target enzyme in its inactive conformation. These conformation-specific scFvs illustrate that stabilization of oxidized PTP1B is an effective strategy to inhibit PTP1B function; it is possible that this approach may be applicable to the protein tyrosine phosphatase (PTP) family as a whole. With this protocol, isolation and characterization of specific scFvs from immune responsive animals should take ~6 weeks. |
| | |
Authors:
|
Aftabul Haque; Nicholas K Tonks |
Related Documents
:
|
9427224 - Peripheral neuropathy associated with igm monoclonal gammopathy: correlations between m... 2410454 - In vitro synthesis of immunoglobulins and autoantibodies by lymphocytes from various bo... 11342254 - Characterization of blood group abo(h)-active gangliosides in type ab erythrocytes and ... 6207204 - A novel myelin-associated glycoprotein defined by a mouse monoclonal antibody. 1945524 - A monoclonal antibody against echinococcus multilocularis em2 antigen. 2140594 - An antibody directed against a peptide encoded by rna complementary to mrna for vasopre... |
Publication Detail:
|
Type: Journal Article; Research Support, N.I.H., Extramural Date: 2012-11-15 |
Journal Detail:
|
Title: Nature protocols Volume: 7 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2012 Dec |
Date Detail:
|
Created Date: 2012-11-16 Completed Date: 2013-04-16 Revised Date: 2013-04-17 |
Medline Journal Info:
|
Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
|
Languages: eng Pagination: 2127-43 Citation Subset: IM |
Affiliation:
|
Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, USA. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Cell Surface Display Techniques
/
methods* Epitopes / metabolism Immunologic Techniques / methods* Oxidation-Reduction Protein Tyrosine Phosphatase, Non-Receptor Type 1 / metabolism* Recombinant Proteins / metabolism* Single-Chain Antibodies / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
|
CA53840/CA/NCI NIH HHS; GM55989/GM/NIGMS NIH HHS; R01 CA053840/CA/NCI NIH HHS; R01 GM055989/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
|
0/Epitopes; 0/Recombinant Proteins; 0/Single-Chain Antibodies; EC 3.1.3.48/Protein Tyrosine Phosphatase, Non-Receptor Type 1 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Exosome-mediated delivery of siRNA in vitro and in vivo.
Next Document: Bifunctional heterometallic Ln(3+)-Gd(3+) (Ln = Eu, Tb) hybrid silica microspheres: luminescence and...