Mapping of linear epitopes recognized by monoclonal antibodies with gene-fragment phage display libraries

1995 ◽  
Vol 249 (4) ◽  
pp. 425-431 ◽  
Author(s):  
Gabriele Petersen ◽  
Danying Song ◽  
Barbara Hügle-Dörr ◽  
Imke Oldenburg ◽  
Ekkehard K. F. Bautz
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Gene Fragment ◽  
Phage Display Libraries ◽  
Linear Epitopes

scholarly journals Generation of Lamprey Monoclonal Antibodies (Lampribodies) Using the Phage Display System

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 868 ◽  
Author(s):  
Khan M. A. Hassan ◽  
John D. Hansen ◽  
Brantley R. Herrin ◽  
Chris T. Amemiya
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Genomic Rearrangements ◽  
Filamentous Phage ◽  
Human Cell Lines ◽  
Display System ◽  
Phage Display Libraries ◽  
Humoral Antibodies ◽  
Leucine Rich Repeats ◽  
Lymphocyte Receptors

The variable lymphocyte receptors (VLRs) consist of leucine rich repeats (LRRs) and comprise the humoral antibodies produced by lampreys and hagfishes. The diversity of the molecules is generated by stepwise genomic rearrangements of LRR cassettes dispersed throughout the VLRB locus. Previously, target-specific monovalent VLRB antibodies were isolated from sea lamprey larvae after immunization with model antigens. Further, the cloned VLR cDNAs from activated lamprey leukocytes were transfected into human cell lines or yeast to select best binders. Here, we expand on the overall utility of the VLRB technology by introducing it into a filamentous phage display system. We first tested the efficacy of isolating phage into which known VLRB molecules were cloned after a series of dilutions. These experiments showed that targeted VLRB clones could easily be recovered even after extensive dilutions (1 to 109). We further utilized the system to isolate target-specific “lampribodies” from phage display libraries from immunized animals and observed an amplification of binders with relative high affinities by competitive binding. The lampribodies can be individually purified and ostensibly utilized for applications for which conventional monoclonal antibodies are employed.

Identification of Major Linear Epitopes on the sp100 Nuclear PBC Autoantigen by the Gene-Fragment Phage-Display Technology

Autoimmunity ◽  
1999 ◽  
Vol 29 (1) ◽  
pp. 33-42 ◽  
Author(s):  
Martin Blüthner ◽  
Cornelia Schäfer ◽  
Carsten Schneider ◽  
Friedlinde A. Bautz
Keyword(s):  
Phage Display ◽  
Gene Fragment ◽  
Phage Display Technology ◽  
Display Technology ◽  
Linear Epitopes

Cloning and expression of human V-genes derived from phage display libraries as fully assembled human anti-TNFα monoclonal antibodies

1997 ◽  
Vol 3 (1) ◽  
pp. 31-43 ◽  
Author(s):  
Stephen M. Mahler ◽  
Chris P. Marquis ◽  
Gary Brown ◽  
Andy Roberts ◽  
Hennie R. Hoogenboom
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Cloning And Expression ◽  
Phage Display Libraries ◽  
V Genes

scholarly journals Developing Recombinant Antibodies by Phage Display Against Infectious Diseases and Toxins for Diagnostics and Therapy

2021 ◽  
Vol 11 ◽  
Author(s):  
Kristian Daniel Ralph Roth ◽  
Esther Veronika Wenzel ◽  
Maximilian Ruschig ◽  
Stephan Steinke ◽  
Nora Langreder ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Infectious Diseases ◽  
Phage Display ◽  
Antibody Fragment ◽  
Recombinant Antibodies ◽  
Phage Display Libraries ◽  
Antibody Phage ◽  
Antibody Phage Display

Antibodies are essential molecules for diagnosis and treatment of diseases caused by pathogens and their toxins. Antibodies were integrated in our medical repertoire against infectious diseases more than hundred years ago by using animal sera to treat tetanus and diphtheria. In these days, most developed therapeutic antibodies target cancer or autoimmune diseases. The COVID-19 pandemic was a reminder about the importance of antibodies for therapy against infectious diseases. While monoclonal antibodies could be generated by hybridoma technology since the 70ies of the former century, nowadays antibody phage display, among other display technologies, is robustly established to discover new human monoclonal antibodies. Phage display is an in vitro technology which confers the potential for generating antibodies from universal libraries against any conceivable molecule of sufficient size and omits the limitations of the immune systems. If convalescent patients or immunized/infected animals are available, it is possible to construct immune phage display libraries to select in vivo affinity-matured antibodies. A further advantage is the availability of the DNA sequence encoding the phage displayed antibody fragment, which is packaged in the phage particles. Therefore, the selected antibody fragments can be rapidly further engineered in any needed antibody format according to the requirements of the final application. In this review, we present an overview of phage display derived recombinant antibodies against bacterial, viral and eukaryotic pathogens, as well as microbial toxins, intended for diagnostic and therapeutic applications.

Mapping of epitopes recognized by autoantibodies with gene-fragment phage display libraries

1996 ◽  
Vol 198 (2) ◽  
pp. 187-198 ◽  
Author(s):  
Martin Blüthner ◽  
Ekkehard K.F. Bautz ◽  
Friedlinde A. Bautz
Keyword(s):  
Phage Display ◽  
Gene Fragment ◽  
Phage Display Libraries

Development of tularemic scFv antibody fragments using phage display

2010 ◽  
Vol 5 (3) ◽  
pp. 310-317
Author(s):  
Klara Kubelkova ◽  
Ales Macela
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Signal Sequence ◽  
Polyclonal Antibodies ◽  
Soluble Form ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Chain ◽  
Bacterial Cell Lysate ◽  
Phage Display Libraries ◽  
Single Fold

AbstractPolyclonal antibodies, as well as monoclonal antibodies are efficacious in providing protective immunity against Francisella tularensis. This study demonstrates the application of phage display libraries for the construction of monoclonal antibodies against F. tularensis. Novel single-chain fragment variable (scFv) antibodies were generated against a whole bacterial lysate of F. tularensis live vaccine strain using the human single fold scFv libraries I (Tomlinson I + J). A total of 20 clones reacted with the bacterial cell lysate. Further, the library contains two clones responsive to recombinant lipoprotein FTT1103Δsignal (F. tularensis subsp. tularensis Schu S4), which was constructed without a signal sequence. These positively-binding scFvs were evaluated by scFv-phage enzyme-linked immunosorbent assay (ELISA). Then, positive scFvs were expressed in a soluble form in Escherichia coli HB2151 and tested for positive scFvs by using scFv-ELISA.

scholarly journals Selection and Characterization of YKL-40-Targeting Monoclonal Antibodies from Human Synthetic Fab Phage Display Libraries

2020 ◽  
Vol 21 (17) ◽  
pp. 6354
Author(s):  
Kyungjae Kang ◽  
Kicheon Kim ◽  
Se-Ra Lee ◽  
Yoonji Kim ◽  
Joo Eon Lee ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Phage Display ◽  
Cancer Therapeutics ◽  
Cell Types ◽  
Migration Assay ◽  
Synthetic Antibody ◽  
Phage Display Libraries ◽  
Anti Cancer

YKL-40, also known as chitinase-3-like 1 (CHI3L1), is a glycoprotein that is expressed and secreted by various cell types, including cancers and macrophages. Due to its implications for and upregulation in a variety of diseases, including inflammatory conditions, fibrotic disorders, and tumor growth, YKL-40 has been considered as a significant therapeutic biomarker. Here, we used a phage display to develop novel monoclonal antibodies (mAbs) targeting human YKL-40 (hYKL-40). Human synthetic antibody phage display libraries were panned against a recombinant hYKL-40 protein, yielding seven unique Fabs (Antigen-binding fragment), of which two Fabs (H1 and H2) were non-aggregating and thermally stable (75.5 °C and 76.5 °C, respectively) and had high apparent affinities (KD = 2.3 nM and 4.0 nM, respectively). Reformatting the Fabs into IgGs (Immunoglobulin Gs) increased their apparent affinities (notably, for H1 and H2, KD = 0.5 nM and 0.3 nM, respectively), presumably due to the effects of avidity, with little change to their non-aggregation property. The six anti-hYKL-40 IgGs were analyzed using a trans-well migration assay in vitro, revealing that three clones (H1, H2, and H4) were notably effective in reducing cell migration from both A549 and H460 lung cancer cell lines. The three clones were further analyzed in an in vivo animal test that assessed their anti-cancer activities, demonstrating that the tumor area and the number of tumor nodules were significantly reduced in the lung tissues treated with H1 (IgG). Given its high affinity and desirable properties, we expect that the H1 anti-hYKL-40 mAb will be a suitable candidate for developing anti-cancer therapeutics.

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