title: Application of streptavidin mass spectrometric immunoassay tips for immunoaffinity based antibody phage display panning.
abstract: Antibody phage display panning involves the enrichment of antibodies against specific targets by affinity. In recent years, several new methods for panning have been introduced to accommodate the growing application of antibody phage display. The present work is concerned with the application of streptavidin mass spectrometry immunoassay (MSIA™) Disposable Automation Research Tips (D.A.R.T's®) for antibody phage display. The system was initially designed to isolate antigens by affinity selection for mass spectrometry analysis. The streptavidin MSIA™ D.A.R.T's® system allows for easy attachment of biotinylated target antigens on the solid surface for presentation to the phage library. As proof-of-concept, a domain antibody library was passed through the tips attached with the Hemolysin E antigen. After binding and washing, the bound phages were eluted via standard acid dissociation and the phages were rescued for subsequent panning rounds. Polyclonal enrichment was observed for three rounds of panning with five monoclonal domain antibodies identified. The proposed method allows for a convenient, rapid and semi-automated alternative to conventional antibody panning strategies.
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: Fisher Scientific (M) Sdn Bhd, No. 3, Jalan Sepadu 25/123, Taman Perindustrian Axis, Seksyen 25, 40400 Shah Alam, Selangor Darul Ehsan, Malaysia.
authors.forename: Lian Wee
authors.initials: LW
authors.lastname: Ler
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: Eugene Boon Beng
authors.initials: EB
authors.lastname: Ong
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.
authors.forename: Gee Jun
authors.initials: GJ
authors.lastname: Tye
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: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000915:Antibody Affinity; D000941:Antigens / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D001435:Bacteriophages / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D019921:Biotinylation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D029968:Escherichia coli Proteins / Q000737:chemistry; D006460:Hemolysin Proteins / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D019151:Peptide Library; D019809:Streptavidin / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000941:Antigens; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; D019151:Peptide Library; C119944:hlyE protein, E coli; D019809:Streptavidin
keywords: Disposable Automation Research Tips (D.A.R.T's®); Domain antibodies; Mass spectrometry immunoassay (MSIA™); Phage display; Semi-automated panning