Cloning of Single-Chain Antibody Variants by Overlap-Extension PCR for Evaluation of Antibody Expression in Transient Gene Expression

2017 ◽  
pp. 57-69 ◽  
Author(s):  
Patrick Mayrhofer ◽  
Renate Kunert
Keyword(s):  
Gene Expression ◽  
Transient Gene Expression ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Antibody Expression ◽  
Overlap Extension Pcr ◽  
Overlap Extension

A Combination Strategy for Construction of Peptide-��2m-H-2Kb Single Chain with Overlap Extension PCR and One-Step Cloning

2016 ◽  
Vol 26 (12) ◽  
pp. 2184-2191 ◽  
Author(s):  
Tao Xu ◽  
Xiaoe Li ◽  
You Wu ◽  
Khawar Ali Shahzad ◽  
Wei Wang ◽  
...  
Keyword(s):  
Single Chain ◽  
Combination Strategy ◽  
Overlap Extension Pcr ◽  
Overlap Extension ◽  
One Step

scholarly journals Human single-chain antibody expression in the hemolymph and fat body of silkworm larvae and pupae using BmNPV bacmids

2009 ◽  
Vol 107 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Motoki Ishikiriyama ◽  
Takuya Nishina ◽  
Tatsuya Kato ◽  
Hiroshi Ueda ◽  
Enoch Y. Park
Keyword(s):  
Fat Body ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Silkworm Larvae ◽  
Antibody Expression

Recombinant antineuraminidase single chain antibody: Expression, characterization, and crystallization in complex with antigen

1993 ◽  
Vol 16 (1) ◽  
pp. 57-63 ◽  
Author(s):  
Robyn L. Malby ◽  
J. Bruce Caldwell ◽  
L. Clem Gruen ◽  
Vincent R. Harley ◽  
Neva Ivancic ◽  
...  
Keyword(s):  
Single Chain ◽  
Single Chain Antibody ◽  
Antibody Expression ◽  
Expression Characterization

scholarly journals Anti-herbicide single-chain antibody expression confers herbicide tolerance in transgenic plants

FEBS Letters ◽  
2003 ◽  
Vol 550 (1-3) ◽  
pp. 179-184 ◽  
Author(s):  
Junichi Eto ◽  
Yoshihito Suzuki ◽  
Hideo Ohkawa ◽  
Isomaro Yamaguchi
Keyword(s):  
Transgenic Plants ◽  
Herbicide Tolerance ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Antibody Expression

Transformation of tobacco plants for single-chain antibody expression via apoplastic and symplasmic routes, and analysis of their susceptibility to stolbur phytoplasma infection

Plant Science ◽  
2005 ◽  
Vol 168 (2) ◽  
pp. 349-358 ◽  
Author(s):  
Sylvie Malembic-Maher ◽  
Fabrice Le Gall ◽  
Jean-Luc Danet ◽  
François Dorlhac de Borne ◽  
Joseph-Marie Bové ◽  
...  
Keyword(s):  
Single Chain ◽  
Single Chain Antibody ◽  
Tobacco Plants ◽  
Antibody Expression ◽  
Phytoplasma Infection ◽  
Stolbur Phytoplasma

scholarly journals A tumor specific single chain antibody dependent gene expression system

Oncogene ◽  
1999 ◽  
Vol 18 (2) ◽  
pp. 559-564 ◽  
Author(s):  
Marie-Noëlle Mary ◽  
Corinne Venot ◽  
Claude Caron de Fromentel ◽  
Laurent Debussche ◽  
Emmanuel Conseiller ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Gene Expression System

scholarly journals Targeted inhibition of HBV gene expression by single-chain antibody mediated small interfering RNA delivery

Hepatology ◽  
2007 ◽  
Vol 46 (1) ◽  
pp. 84-94 ◽  
Author(s):  
Wei-Hong Wen ◽  
Jia-Yun Liu ◽  
Wei-Jun Qin ◽  
Jing Zhao ◽  
Tao Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Interfering Rna ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Targeted Inhibition ◽  
Interfering Rna

A New Disulfide-Linked Dimer of a Single-Chain Antibody Fragment Against Human CD47 Induces Apoptosis in Lymphoid Malignant Cells In Vitro and In Vivo Via the HIF-1α Pathway: A Possible Novel Therapeutic Agent for B-CLL.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2097-2097
Author(s):  
Morihiko Sagawa ◽  
Takatsune Shimizu ◽  
Mingji Xian ◽  
Aki Chizuka ◽  
Naoshi Fukushima ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression Profiling ◽  
Therapeutic Agent ◽  
Antibody Fragment ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Lymphoid Malignancies ◽  
Induced Apoptosis ◽  
Single Chain Antibody Fragment ◽  
Novel Therapeutic

Abstract CD47 belongs to the immunoglobulin superfamily, and is expressed as a 50 kDa cell surface antigen in a wide variety of tissues. CD47 is associated with integrins of β1, β2, and β3, and serves as both a receptor for thrombospondin (TSP) and a ligand for the transmembrane signal regulatory protein SIRP-α . CD47 has a number of different functions, including platelet activation, cell motility, leukocyte adhesion, migration and phagocytosis. Recently, it was reported that the ligation of CD47 rapidly induces cell death in T-cells and chronic lymphocytic leukemic B cells (B-CLL) in a caspase-independent manner (Nat Med, 1999;5:1277–1284). These findings indicate that CD47 ligation may have potential as a new therapeutic approach for lymphoid malignancies. B-CLL is the most common hematological malignancy in Western countries. Although new therapeutic agents such as fludarabine and 2-chlorodeoxyadenosine have been introduced in the clinic, B-CLL is not considered curable, and thus there is an immediate need for new, effective drugs. In an attempt to establish a novel therapeutic agent for B-CLL, we first generated a murine monoclonal antibody against an extracellular domain of human CD47 (designated MABL). Soluble MABL (10μg/ml) with goat anti-mouse IgG (GAM) induced apoptosis of CD47-positive CCRF-CEM and MOLT-4 cells. However, because MABL caused hemoagglutination, we created a disulfide-stabilized dimer of a single-chain antibody fragment of MABL (S-S diabody) to get rid of this adverse effect. The S-S diabody did not cause hemoagglutination. Treatment with the S-S diabody (0–0.1 μg/ml for 0–24 hours) alone was fully active in inducing apoptosis of primary samples from patients with B-CLL and CD47-positive lymphoid cells (MOLT-4 and JOK-1) in a dose- and time-dependent manner. In addition, administration of the S-S diabody (30 mg/kg for 5 days) significantly prolonged the survival of SCID mice inoculated with JOK-1 cells compared with that of the control mice (median survival 21 days vs. more than 30 days, p<0.05). These results demonstrate that the S-S diabody induced the ligation of CD47 more efficiently than original MABL without causing hemoagglutination. Transmission electron microscopic analysis showed that the S-S diabody induced typical morphological changes of apoptosis as well as cell-cell adhesion in MOLT-4 cells. To address the molecular mechanism of apoptosis by the ligation of CD47 with the S-S diabody, we performed gene expression profiling analysis. Gene expression profiling with Affimetrix gene chips of MOLT-4 cells demonstrated 54 genes, including HIF-1a and its down-stream genes (RTP801, EGR-1, BNIP3, and VEGF), that were upregulated by the treatment with S-S diabody. Moreover, knockdown of these molecules by siRNA partially repressed S-S diabody-induced apoptosis in MOLT-4 cells. These results suggest that induction of apoptosis by CD47 ligation may be mediated through various HIF-1α pathways. In conclusion, CD47 will be a molecular target for the treatment of lymphoid malignancies. In addition, the newly established disulfide-stabilized dimer of a single-chain antibody fragment of MABL (S-S diabody) may have potential as a novel therapeutic agent for the treatment of B-CLL and other incurable lymphoid malignancies.

scholarly journals Genome-wide analysis of a G-quadruplex-specific single-chain antibody that regulates gene expression

2009 ◽  
Vol 37 (20) ◽  
pp. 6716-6722 ◽  
Author(s):  
Himesh Fernando ◽  
Sven Sewitz ◽  
Jeremy Darot ◽  
Simon Tavaré ◽  
Julian Leon Huppert ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Chain ◽  
Single Chain Antibody ◽  
Genome Wide Analysis ◽  
Genome Wide ◽  
G Quadruplex
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