Utilization of multi-immunization and multiple selection strategies for isolation of hapten-specific antibodies from recombinant antibody phage display libraries

Antti Tullila, Tarja K. Nevanen

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)

Abstract

Phage display technology provides a powerful tool for the development of novel recombinant antibodies. In this work, we optimized and streamlined the recombinant antibody discovery process for haptens as an example. A multi-immunization approach was used in order to avoid the need for construction of multiple antibody libraries. Selection methods were developed to utilize the full potential of the recombinant antibody library by applying four different elution conditions simultaneously. High-throughput immunoassays were used to analyse the binding properties of the individual antibody clones. Different carrier proteins were used in the immunization, selection, and screening phases to avoid enrichment of the antibodies for the carrier protein epitopes. Novel recombinant antibodies against mycophenolic acid and ochratoxin A, with affinities up to 39 nM and 34 nM, respectively, were isolated from a multi-immunized fragment antigen-binding (Fab) library.
Original languageEnglish
Article number1169
JournalInternational Journal of Molecular Sciences
Volume18
Issue number6
DOIs
Publication statusPublished - 1 Jun 2017
MoE publication typeA1 Journal article-refereed

Fingerprint

Immunization
Bacteriophages
Haptens
antibodies
Antibodies
Libraries
isolation
Display devices
Carrier Proteins
Mycophenolic Acid
proteins
Epitopes
immunoassay
elution
antigens
Antigens
Immunoassay
affinity
Screening
screening

Keywords

  • fragment antigen-binding (Fab)
  • hapten
  • high-throughput
  • mycotoxin
  • phage display
  • recombinant antibody

Cite this

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