Phage Display Vectors for the In Vitro Generation of Human Antibody Fragments

2005 ◽  
pp. 71-96 ◽  
Author(s):  
Michael Hust ◽  
Stefan Dübel
Keyword(s):  
Phage Display ◽  
Antibody Fragments ◽  
Human Antibody

scholarly journals A novel and effective approach to generate germline-like monoclonal antibodies by integration of phage and mammalian cell display platforms

2021 ◽  
Author(s):  
Yu-jia Jin ◽  
Diao Yu ◽  
Xiao-long Tian ◽  
Hui-xian Li ◽  
Xiao-chao Zhou ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Phage Display ◽  
Mammalian Cell ◽  
Phage Display Library ◽  
Antibody Fragments ◽  
Human Antibody ◽  
Phage Display Technology ◽  
Mammalian Cell Display

AbstractPhage display technology allows for rapid selection of antibodies from the large repertoire of human antibody fragments displayed on phages. However, antibody fragments should be converted to IgG for biological characterizations and affinity of antibodies obtained from phage display library is frequently not sufficient for efficient use in clinical settings. Here, we describe a new approach that combines phage and mammalian cell display, enabling simultaneous affinity screening of full-length IgG antibodies. Using this strategy, we successfully obtained a novel germline-like anti-TIM-3 monoclonal antibody named m101, which was revealed to be a potent anti-TIM-3 therapeutic monoclonal antibody via in vitro and in vivo experiments, indicating its effectiveness and power. Thus, this platform can help develop new monoclonal antibody therapeutics with high affinity and low immunogenicity.

1278 Targeting anti-fibrogenic therapeutics to hepatic stellate cells — generating human antibody fragments by phage display

Hepatology ◽  
2003 ◽  
Vol 38 ◽  
pp. 775-775
Author(s):  
L ELRICK ◽  
V LEEL ◽  
C MAREK ◽  
N KORUTH ◽  
K CHARLTON ◽  
...  
Keyword(s):  
Phage Display ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Antibody Fragments ◽  
Human Antibody

scholarly journals Generation of human antibody fragments recognizing distinct epitopes of the nucleocapsid (N) SARS-CoV protein using a phage display approach

2005 ◽  
Vol 5 (1) ◽  
Author(s):  
Michela Flego ◽  
Paola Di Bonito ◽  
Alessandro Ascione ◽  
Silvia Zamboni ◽  
Alessandra Carattoli ◽  
...  
Keyword(s):  
Phage Display ◽  
Antibody Fragments ◽  
Human Antibody

Identification of Dengue-Specific Human Antibody Fragments Using Phage Display

2014 ◽  
pp. 161-173 ◽  
Author(s):  
Moon Y. F. Tay ◽  
Chin Chin Lee ◽  
Subhash G. Vasudevan ◽  
Nicole J. Moreland
Keyword(s):  
Phage Display ◽  
Antibody Fragments ◽  
Human Antibody

Functional Analysis of Phage Display Selected Single-Chain Variable Antibody Fragments (scFvs) Specific for Anti-FVIII Antibodies

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1499-1499
Author(s):  
Kerstin Brettschneider ◽  
Anja Naumann ◽  
Sonja Neimanis ◽  
Joerg Kahle ◽  
Christine Heller ◽  
...  
Keyword(s):  
Phage Display ◽  
Hemophilia A ◽  
Antibody Fragments ◽  
Dependent Manner ◽  
Single Chain ◽  
Concentration Dependent Manner ◽  
Fviii Activity ◽  
Inhibitory Antibodies

Abstract The development of inhibitory antibodies against coagulation factor VIII (FVIII) is currently the most serious complication for hemophilia A patients that undergo FVIII replacement therapy. In addition, non-hemophilia A patients can spontaneously develop inhibitory auto-antibodies to FVIII, which results in acquired haemophilia A. The control of the allo- or autoimmune response to FVIII apparently includes the elicitation of anti-idiotypic antibodies. In this study the capacity of anti-idiotypic single-chain variable antibody fragments (scFvs) for neutralization of inhibitory anti-FVIII antibodies (FVIII inhibitors) was evaluated in vitro and in vivo. Anti-idiotypic scFvs were selected from phage-displayed libraries against murine monoclonal FVIII-specific inhibitors. As the majority of inhibitory antibodies is directed against the A2 or C2 domain of FVIII, strongly inhibitory A2- and C2-specific antibodies served as targets. Selected scFvs were expressed as scFv-Fc fusion proteins. Analysis of the scFv-Fcs by ELISA confirmed the specific binding to the cognate targets and binding studies via surface plasmon resonance revealed high affinities within the nanomolar range. Further characterization showed that binding of inhibitors to immobilized FVIII was blocked by specific scFv-Fcs in vitro. The ability of scFv-Fcs to neutralize their corresponding inhibitors was analyzed in a functional clotting assay. By adding scFv-Fcs to plasma spiked with inhibitors, FVIII activity was restored to 80% in a concentration dependent manner. FVIII knockout mice served as model organism for testing the capacity of scFv-Fcs to restore coagulation in vivo. Subsequent injection of FVIII following the injection of the inhibitors resulted in a largely reduced FVIII activity. However, FVIII activity was recovered in a concentration dependent manner by adding cognate anti-idiotypes. The scFv-Fcs were either preincubated with the corresponding inhibitor or added to the FVIII mixture without preincubation. The latter represents an adaption to a therapeutic setting. In conclusion, phage display selected anti-idiotypic scFvs are able to bind and effectively neutralize their target inhibitors in vitro and in vivo. Based on these promising results the potential of anti-idiotypic scFvs for the development of specific cell based immunotherapies for hemophilia A patients with inhibitors is currently under investigation. Disclosures No relevant conflicts of interest to declare.

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