scholarly journals High Affinity Maturated Human Antibodies from Naïve and Synthetic Antibody Repertoires

2018 ◽  
Author(s):  
Chia Chiu Lim ◽  
Yee Siew Choong ◽  
Theam Soon Lim
Keyword(s):  
Synthetic Antibody ◽  
Human Antibodies ◽  
High Affinity ◽  
Antibody Repertoires

scholarly journals Common light chain chickens produce human antibodies of high affinity and broad epitope coverage for the engineering of bispecifics

mAbs ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 1862451
Author(s):  
Kathryn H. Ching ◽  
Kimberley Berg ◽  
Kevin Reynolds ◽  
Darlene Pedersen ◽  
Alba Macias ◽  
...  
Keyword(s):  
Light Chain ◽  
Human Antibodies ◽  
High Affinity

scholarly journals Recognition of an α-helical hairpin in P22 large terminase by a synthetic antibody fragment

2020 ◽  
Vol 76 (9) ◽  
pp. 876-888
Author(s):  
Ravi K. Lokareddy ◽  
Ying-Hui Ko ◽  
Nathaniel Hong ◽  
Steven G. Doll ◽  
Marcin Paduch ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Conformational Changes ◽  
Recombinant Antibody ◽  
Antibody Fragment ◽  
Genome Packaging ◽  
Synthetic Antibody ◽  
High Affinity ◽  
N Terminus ◽  
Helical Hairpin ◽  
Accessible Surface

The genome-packaging motor of tailed bacteriophages and herpesviruses is a multisubunit protein complex formed by several copies of a large (TerL) and a small (TerS) terminase subunit. The motor assembles transiently at the portal protein vertex of an empty precursor capsid to power the energy-dependent packaging of viral DNA. Both the ATPase and nuclease activities associated with genome packaging reside in TerL. Structural studies of TerL from bacteriophage P22 have been hindered by the conformational flexibility of this enzyme and its susceptibility to proteolysis. Here, an unbiased, synthetic phage-display Fab library was screened and a panel of high-affinity Fabs against P22 TerL were identified. This led to the discovery of a recombinant antibody fragment, Fab4, that binds a 33-amino-acid α-helical hairpin at the N-terminus of TerL with an equilibrium dissociation constant K d of 71.5 nM. A 1.51 Å resolution crystal structure of Fab4 bound to the TerL epitope (TLE) together with a 1.15 Å resolution crystal structure of the unliganded Fab4, which is the highest resolution ever achieved for a Fab, elucidate the principles governing the recognition of this novel helical epitope. TLE adopts two different conformations in the asymmetric unit and buries as much as 1250 Å2 of solvent-accessible surface in Fab4. TLE recognition is primarily mediated by conformational changes in the third complementarity-determining region of the Fab4 heavy chain (CDR H3) that take place upon epitope binding. It is demonstrated that TLE can be introduced genetically at the N-terminus of a target protein, where it retains high-affinity binding to Fab4.

scholarly journals High-affinity self-reactive human antibodies by design and selection: targeting the integrin ligand binding site.

1993 ◽  
Vol 90 (21) ◽  
pp. 10003-10007 ◽  
Author(s):  
C. F. Barbas ◽  
L. R. Languino ◽  
J. W. Smith
Keyword(s):  
Ligand Binding ◽  
Binding Site ◽  
Ligand Binding Site ◽  
Human Antibodies ◽  
High Affinity ◽  
Integrin Ligand

Cloning, sequencing and characterization of high affinity human antibodies against HCV NS3-protease/helicase by phage display

2001 ◽  
Vol 34 ◽  
pp. 121
Author(s):  
T. Heintges ◽  
K. Kraszucka ◽  
A. Erhardt ◽  
D. Haussinger
Keyword(s):  
Phage Display ◽  
Human Antibodies ◽  
High Affinity ◽  
Ns3 Protease

scholarly journals Affinity-engineered human antibodies detect celiac disease gluten pMHC complexes and inhibit T-cell activation

2019 ◽  
Author(s):  
Rahel Frick ◽  
Lene S. Høydahl ◽  
Ina Hodnebrug ◽  
Shraddha Kumari ◽  
Grete Berntsen ◽  
...  
Keyword(s):  
T Cells ◽  
Celiac Disease ◽  
T Cell ◽  
Antigen Presentation ◽  
T Cell Activation ◽  
Cell Activation ◽  
Plasma Cells ◽  
Human Antibodies ◽  
High Affinity

AbstractAntibodies specific for antigenic peptides bound to major histocompatibility complex (MHC) molecules are valuable tools for studies of antigen presentation. Such T-cell receptor (TCR)-like antibodies may also have therapeutic potential in human disease due to their ability to target disease-associated antigens with high specificity. We previously generated celiac disease (CeD) relevant TCR-like antibodies that recognize the prevalent gluten epitope DQ2.5-glia-α1a in complex with HLA-DQ2.5. Here, we report on second-generation high-affinity antibodies towards this epitope as well as a panel of novel TCR-like antibodies to another immunodominant gliadin epitope, DQ2.5-glia-α2. The strategy for affinity engineering was based on Rosetta modeling combined with pIX phage display and is applicable to similar protein engineering efforts. We isolated picomolar affinity binders and validated them in Fab and IgG format. Flow cytometry experiments with CeD biopsy material confirm the unique disease specificity of these TCR-like antibodies and reinforce the notion that B cells and plasma cells have a dominant role in gluten antigen presentation in the inflamed CeD gut. Further, the lead candidate 3.C11 potently inhibited CD4+ T-cell activation and proliferation in vitro in an HLA and epitope specific manner, pointing to a potential for targeted disease interception without compromising systemic immunity.Significance StatementConsumption of gluten-containing food drives celiac disease in genetically predisposed individuals. The underlying disease mechanism is not fully understood, but it is strictly dependent on activation of pathogenic T cells. We have engineered high-affinity human antibodies recognizing the T-cell target HLA-DQ2.5 in complex with gluten epitopes and studied cell-specific antigen presentation in patients, which shows that plasma cells and not dendritic cells dominate the inflamed tissue. The only available treatment is lifelong adherence to a gluten-free diet, which is difficult and not effective in all cases. We show that at least one of our antibodies can specifically inhibit activation of pathogenic T-cells in vitro and therefore shows promise for therapy.

11 Isolating High Affinity Human Antibodies from Phage Repertoires

2013 ◽  
Author(s):  
Kevin FitzGerald ◽  
David Chiswell ◽  
John Earnshaw ◽  
Rodger Smith ◽  
John Kenten ◽  
...  
Keyword(s):  
Human Antibodies ◽  
High Affinity
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