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Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies

As a novel alternative to established surface display or combinatorial chemistry approaches for the discovery of therapeutic peptides, we present a method for the isolation of small, cysteine-rich domains from bovine antibody ultralong complementarity-determining regions (CDRs). We show for the firs...

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Autores principales: Macpherson, Alex, Scott-Tucker, Anthony, Spiliotopoulos, Anastasios, Simpson, Catherine, Staniforth, Justin, Hold, Adam, Snowden, James, Manning, Leah, van den Elsen, Jean, Lawson, Alastair D. G.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498065/
https://www.ncbi.nlm.nih.gov/pubmed/32886672
http://dx.doi.org/10.1371/journal.pbio.3000821
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author Macpherson, Alex
Scott-Tucker, Anthony
Spiliotopoulos, Anastasios
Simpson, Catherine
Staniforth, Justin
Hold, Adam
Snowden, James
Manning, Leah
van den Elsen, Jean
Lawson, Alastair D. G.
author_facet Macpherson, Alex
Scott-Tucker, Anthony
Spiliotopoulos, Anastasios
Simpson, Catherine
Staniforth, Justin
Hold, Adam
Snowden, James
Manning, Leah
van den Elsen, Jean
Lawson, Alastair D. G.
author_sort Macpherson, Alex
collection PubMed
description As a novel alternative to established surface display or combinatorial chemistry approaches for the discovery of therapeutic peptides, we present a method for the isolation of small, cysteine-rich domains from bovine antibody ultralong complementarity-determining regions (CDRs). We show for the first time that isolated bovine antibody knob domains can function as autonomous entities by binding antigen outside the confines of the antibody scaffold. This yields antibody fragments so small as to be considered peptides, each stabilised by an intricate, bespoke arrangement of disulphide bonds. For drug discovery, cow immunisations harness the immune system to generate knob domains with affinities in the picomolar to low nanomolar range, orders of magnitude higher than unoptimized peptides from naïve library screening. Using this approach, knob domain peptides that tightly bound Complement component C5 were obtained, at scale, using conventional antibody discovery and peptide purification techniques.
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spelling pubmed-74980652020-09-24 Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies Macpherson, Alex Scott-Tucker, Anthony Spiliotopoulos, Anastasios Simpson, Catherine Staniforth, Justin Hold, Adam Snowden, James Manning, Leah van den Elsen, Jean Lawson, Alastair D. G. PLoS Biol Methods and Resources As a novel alternative to established surface display or combinatorial chemistry approaches for the discovery of therapeutic peptides, we present a method for the isolation of small, cysteine-rich domains from bovine antibody ultralong complementarity-determining regions (CDRs). We show for the first time that isolated bovine antibody knob domains can function as autonomous entities by binding antigen outside the confines of the antibody scaffold. This yields antibody fragments so small as to be considered peptides, each stabilised by an intricate, bespoke arrangement of disulphide bonds. For drug discovery, cow immunisations harness the immune system to generate knob domains with affinities in the picomolar to low nanomolar range, orders of magnitude higher than unoptimized peptides from naïve library screening. Using this approach, knob domain peptides that tightly bound Complement component C5 were obtained, at scale, using conventional antibody discovery and peptide purification techniques. Public Library of Science 2020-09-04 /pmc/articles/PMC7498065/ /pubmed/32886672 http://dx.doi.org/10.1371/journal.pbio.3000821 Text en © 2020 Macpherson et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Methods and Resources
Macpherson, Alex
Scott-Tucker, Anthony
Spiliotopoulos, Anastasios
Simpson, Catherine
Staniforth, Justin
Hold, Adam
Snowden, James
Manning, Leah
van den Elsen, Jean
Lawson, Alastair D. G.
Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title_full Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title_fullStr Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title_full_unstemmed Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title_short Isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
title_sort isolation of antigen-specific, disulphide-rich knob domain peptides from bovine antibodies
topic Methods and Resources
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498065/
https://www.ncbi.nlm.nih.gov/pubmed/32886672
http://dx.doi.org/10.1371/journal.pbio.3000821
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