Generation and isolation of target-specific single-domain antibodies from shark immune repertoires

Mischa Roland Müller, Ronan O'Dwyer, Marina Kovaleva, Fiona Rudkin, Helen Dooley, Caroline Barelle

Research output: Chapter in Book/Report/Conference proceedingChapter

21 Citations (Scopus)

Abstract

The drive to exploit novel targets and biological pathways has lead to the expansion of classical antibody research into innovative fragment adaptations and novel scaffolds. The hope being that alternative or cryptic epitopes may be targeted, tissue inaccessibility may be overcome, and easier engineering options will facilitate multivalent, multi-targeting approaches. To this end, we have been isolating shark single domains to gain a greater understanding of their potential as therapeutic agents. Their unique shape, small size, inherent stability, and simple molecular architecture make them attractive candidates from a drug discovery perspective. Here we describe protocols to capture the immune repertoire of an immunized shark species and to build and select via phage-display target-specific IgNAR variable domains (VNARs).
Original languageEnglish
Title of host publicationMethods in Molecular Biology
Subtitle of host publicationAntibody Engineering
EditorsPatrick Chames
PublisherHumana Press
Pages177-194
Number of pages18
Volume907
ISBN (Electronic)978-1-61779-974-7
ISBN (Print)978-1-61779-973-0
DOIs
Publication statusPublished - 1 Oct 2012

Publication series

NameMethods and Protocols
PublisherHumana Press
ISSN (Print)1064-3745

Fingerprint

Single-Domain Antibodies
Sharks
Drug Discovery
Bacteriophages
Epitopes
Antibodies
Research
Therapeutics

Keywords

  • shark
  • IgNAR
  • single binding domain
  • phage display
  • immunization
  • immune repertoire
  • selection
  • screening

Cite this

Müller, M. R., O'Dwyer, R., Kovaleva, M., Rudkin, F., Dooley, H., & Barelle, C. (2012). Generation and isolation of target-specific single-domain antibodies from shark immune repertoires. In P. Chames (Ed.), Methods in Molecular Biology: Antibody Engineering (Vol. 907, pp. 177-194). (Methods and Protocols). Humana Press. https://doi.org/10.1007/978-1-61779-974-7_9

Generation and isolation of target-specific single-domain antibodies from shark immune repertoires. / Müller, Mischa Roland; O'Dwyer, Ronan; Kovaleva, Marina; Rudkin, Fiona; Dooley, Helen; Barelle, Caroline.

Methods in Molecular Biology: Antibody Engineering. ed. / Patrick Chames. Vol. 907 Humana Press, 2012. p. 177-194 (Methods and Protocols).

Research output: Chapter in Book/Report/Conference proceedingChapter

Müller, MR, O'Dwyer, R, Kovaleva, M, Rudkin, F, Dooley, H & Barelle, C 2012, Generation and isolation of target-specific single-domain antibodies from shark immune repertoires. in P Chames (ed.), Methods in Molecular Biology: Antibody Engineering. vol. 907, Methods and Protocols, Humana Press, pp. 177-194. https://doi.org/10.1007/978-1-61779-974-7_9
Müller MR, O'Dwyer R, Kovaleva M, Rudkin F, Dooley H, Barelle C. Generation and isolation of target-specific single-domain antibodies from shark immune repertoires. In Chames P, editor, Methods in Molecular Biology: Antibody Engineering. Vol. 907. Humana Press. 2012. p. 177-194. (Methods and Protocols). https://doi.org/10.1007/978-1-61779-974-7_9
Müller, Mischa Roland ; O'Dwyer, Ronan ; Kovaleva, Marina ; Rudkin, Fiona ; Dooley, Helen ; Barelle, Caroline. / Generation and isolation of target-specific single-domain antibodies from shark immune repertoires. Methods in Molecular Biology: Antibody Engineering. editor / Patrick Chames. Vol. 907 Humana Press, 2012. pp. 177-194 (Methods and Protocols).
@inbook{8dddd71fbb85448088683f2f471a881e,
title = "Generation and isolation of target-specific single-domain antibodies from shark immune repertoires",
abstract = "The drive to exploit novel targets and biological pathways has lead to the expansion of classical antibody research into innovative fragment adaptations and novel scaffolds. The hope being that alternative or cryptic epitopes may be targeted, tissue inaccessibility may be overcome, and easier engineering options will facilitate multivalent, multi-targeting approaches. To this end, we have been isolating shark single domains to gain a greater understanding of their potential as therapeutic agents. Their unique shape, small size, inherent stability, and simple molecular architecture make them attractive candidates from a drug discovery perspective. Here we describe protocols to capture the immune repertoire of an immunized shark species and to build and select via phage-display target-specific IgNAR variable domains (VNARs).",
keywords = "shark, IgNAR, single binding domain, phage display, immunization, immune repertoire, selection, screening",
author = "M{\"u}ller, {Mischa Roland} and Ronan O'Dwyer and Marina Kovaleva and Fiona Rudkin and Helen Dooley and Caroline Barelle",
year = "2012",
month = "10",
day = "1",
doi = "10.1007/978-1-61779-974-7_9",
language = "English",
isbn = "978-1-61779-973-0",
volume = "907",
series = "Methods and Protocols",
publisher = "Humana Press",
pages = "177--194",
editor = "Patrick Chames",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Generation and isolation of target-specific single-domain antibodies from shark immune repertoires

AU - Müller, Mischa Roland

AU - O'Dwyer, Ronan

AU - Kovaleva, Marina

AU - Rudkin, Fiona

AU - Dooley, Helen

AU - Barelle, Caroline

PY - 2012/10/1

Y1 - 2012/10/1

N2 - The drive to exploit novel targets and biological pathways has lead to the expansion of classical antibody research into innovative fragment adaptations and novel scaffolds. The hope being that alternative or cryptic epitopes may be targeted, tissue inaccessibility may be overcome, and easier engineering options will facilitate multivalent, multi-targeting approaches. To this end, we have been isolating shark single domains to gain a greater understanding of their potential as therapeutic agents. Their unique shape, small size, inherent stability, and simple molecular architecture make them attractive candidates from a drug discovery perspective. Here we describe protocols to capture the immune repertoire of an immunized shark species and to build and select via phage-display target-specific IgNAR variable domains (VNARs).

AB - The drive to exploit novel targets and biological pathways has lead to the expansion of classical antibody research into innovative fragment adaptations and novel scaffolds. The hope being that alternative or cryptic epitopes may be targeted, tissue inaccessibility may be overcome, and easier engineering options will facilitate multivalent, multi-targeting approaches. To this end, we have been isolating shark single domains to gain a greater understanding of their potential as therapeutic agents. Their unique shape, small size, inherent stability, and simple molecular architecture make them attractive candidates from a drug discovery perspective. Here we describe protocols to capture the immune repertoire of an immunized shark species and to build and select via phage-display target-specific IgNAR variable domains (VNARs).

KW - shark

KW - IgNAR

KW - single binding domain

KW - phage display

KW - immunization

KW - immune repertoire

KW - selection

KW - screening

U2 - 10.1007/978-1-61779-974-7_9

DO - 10.1007/978-1-61779-974-7_9

M3 - Chapter

SN - 978-1-61779-973-0

VL - 907

T3 - Methods and Protocols

SP - 177

EP - 194

BT - Methods in Molecular Biology

A2 - Chames, Patrick

PB - Humana Press

ER -