The general idea of this study was to develop candidate specific IgY antibodies for an oral therapy targeting the ESBL-producing gram negative bacteria. As the family of ESBLs constantly grows and there is lack of their clear classification in the literature, the specific aim was to build a proof of concept study based on the parental enzyme ß-lactamase TEM-1 to investigate different specific IgYs strategies to inhibit the growth of TEM-1 producing E.coli.
This research included a bioinformatic analysis of the TEM-1 structure in the context of TEM-derivative ESBLs. Then, two IgY strategies were designed to target the ß-lactamase TEM-1-producing E.coli (BW25113 ΔbamBΔtolC) with IgYs: as a complement to antibiotics (IgYs against the enzyme TEM-1 used in combination with ampicillin) and as an alternative to antibiotics (IgYs against the bacteria TEM-1-producing E.coli without the addition of ampicillin). A good inhibitory effect of (a)TIgY, (a)p2IgY (in the presence of ampicillin) and eIgY, hIgY (without the ampicillin) on TEM-1-producing E.coli was observed in vitro. Moreover, they had the typical configuration of avian antibodies and were highly specific to their antigens.
This study presents a model system to develop specific IgYs against a therapeutic target of interest. The activity of these IgYs complementary or alternatively to antibiotics should be further investigated in vivo in an animal infectious model. IgYs developed in this study might also be good candidates for further investigation as a broad-spectrum treatment against a variety of ESBL-producing E.coli. The aTIgY which was developed against the whole TEM-1 might also target its derivatives, as they have similar 3D structure with single amino acids mutations in the sequence. The ap2IgY was generated against catalytic and conservative residues, characteristic for the whole class A of ß-lactamases, thus it might target also the active site of ESBL-s from this class. The strategy used to generate eIgY and hIgY was efficient and IgYs could be generated directly against ESBL-producing bacteria.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:21529 |
| Date | 20 June 2018 |
| Creators | Zajac, Julia Dominika |
| Contributors | Horn, Friedemann, Rodloff, Arne C., Sack, Ulrich, Universität Leipzig |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:doctoralThesis, info:eu-repo/semantics/doctoralThesis, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
Page generated in 0.0019 seconds