title: Generation of a naïve human single chain variable fragment (scFv) library for the identification of monoclonal scFv against Salmonella Typhi Hemolysin E antigen.
abstract: Antibody phage display is a useful tool for the isolation and identification of monoclonal antibodies. Naive antibody libraries are able to overcome the limitations associated with the traditional hybridoma method for monoclonal antibody generation. Antibody phage display is also a preferred method for antibody generation against toxins as it does not suffer from toxicity mediated complications. Here, we describe a naïve multi ethnic scFv antibody library generated via two-step cloning with an estimated diversity of 2 × 10(9). The antibody library was used to screen for monoclonal antibodies against Hemolysin E antigen, a pore forming toxin produced by Salmonella enterica serovar Typhi. A soluble monoclonal scFv antibody against the HlyE toxin (IgM scFv D7 anti-hlyE) was isolated from the library. This shows the value of the naïve library to generate antibodies against toxin targets in addition to the potential use of the library to isolate antibodies against other immunogenic targets.
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
authors.forename: Bee Nar
authors.initials: BN
authors.lastname: Lim
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
authors.forename: Chai Fung
authors.initials: CF
authors.lastname: Chin
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
authors.forename: Yee Siew
authors.initials: YS
authors.lastname: Choong
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia.
authors.forename: Asma
authors.initials: A
authors.lastname: Ismail
authors.affiliation: Institute for Research in Molecular Medicine, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia. Electronic address: theamsoon@usm.my.
authors.forename: Theam Soon
authors.initials: TS
authors.lastname: Lim
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins / Q000276:immunology*; D006460:Hemolysin Proteins / Q000276:immunology*; D006801:Humans; D019151:Peptide Library; D012485:Salmonella typhi / Q000235:genetics / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000276:immunology* / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; D019151:Peptide Library; D057127:Single-Chain Antibodies; C119944:hlyE protein, E coli
keywords: Antibody phage display; Hemolysin E; Monoclonal antibodies; Naïve library; Salmonella enterica serovar Typhi; scFv