Evaluation of a nanobody phage display library constructed from a Brucella-immunised camel.

TitleEvaluation of a nanobody phage display library constructed from a Brucella-immunised camel.
Publication TypeJournal Article
Year of Publication2011
AuthorsAbbady, A. Q., A. Al-Mariri, M. Zarkawi, A. Al-Assad, and S. Muyldermans
JournalVet Immunol Immunopathol
Volume142
Issue1-2
Pagination49-56
Date Published2011 Jul 15
ISSN1873-2534
KeywordsAnimals, Brucella melitensis, Brucellosis, Camels, Enzyme-Linked Immunosorbent Assay, Female, Immunity, Humoral, Immunization, Immunoglobulin G, Male, Peptide Library, Single-Chain Antibodies
Abstract

Brucella are invasive gram-negative bacteria that multiply and survive within eukaryotic cells causing brucellosis. Syrian (and Middle East) health and economy sectors are still affected by this disease causing a serious national problem that needs to be solved. Here, a strategy was developed to introduce a new generation of binders, known as Nanobodies (Nbs) in our combat against Brucella. These Nbs, recombinant single-domain variable fragments derived from camelid heavy-chain antibodies are very stable and highly soluble, making them a useful tool in numerous biotechnological and medical applications. In this work and without having access to purified antigens (Ags), a camel was immunised successfully with heat-killed Brucella melitensis strain Riv1 as demonstrated by the high titer of Ag-specific heavy-chain antibodies in the serum. Lymphocytes of the immunised camel were isolated and their Nb genes were cloned in a relatively large library of 10(8) individual transformants, of which 81% contained an insert with the proper size of a Nb gene. Phage display expression of the Nbs from this library and pannings on the Brucella lysate resulted in a clear enrichment of three distinct Nb-displaying phages (phage-Nbs), referred to as NbBruc01, 02 and 03, with specificity for Brucella. Producing these binders in a pure, soluble form, as well as identifying their specific targets, which are likely to be immunodominant Ags in Brucella, is expected to open wide perspectives for following the vaccination, diagnosis and treatment of brucellosis.

DOI10.1016/j.vetimm.2011.04.004
Alternate JournalVet. Immunol. Immunopathol.
PubMed ID21592585