title: A Recombinant Secondary Antibody Mimic as a Target-specific Signal Amplifier and an Antibody Immobilizer in Immunoassays.
abstract: We construct a novel recombinant secondary antibody mimic, GST-ABD, which can bind to the Fc regions of target-bound primary antibodies and acquire multiple HRPs simultaneously. We produce it in tenth of mg quantities with a bacterial overexpression system and simple purification procedures, significantly reducing the manufacturing cost and time without the use of animals. GST-ABD is effectively conjugated with 3 HRPs per molecule on an average and selectively bind to the Fc region of primary antibodies derived from three different species (mouse, rabbit, and rat). HRP-conjugated GST-ABD (HRP-GST-ABD) is successfully used as an alternative to secondary antibodies to amplify target-specific signals in both ELISA and immunohistochemistry regardless of the target molecules and origin of primary antibodies used. GST-ABD also successfully serves as an anchoring adaptor on the surface of GSH-coated plates for immobilizing antigen-capturing antibodies in an orientation-controlled manner for sandwich-type indirect ELISA through simple molecular recognition without any complicated chemical modification.
authors.affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798, Korea.
authors.forename: Junseon
authors.initials: J
authors.lastname: Min
authors.affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798, Korea.
authors.forename: Eun Kyung
authors.initials: EK
authors.lastname: Song
authors.affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798, Korea.
authors.forename: Hansol
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Seoul National University, Seoul, 151-747, Korea.
authors.forename: Kyoung Taek
authors.initials: KT
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798, Korea.
authors.forename: Tae Joo
authors.initials: TJ
authors.lastname: Park
authors.affiliation: Department of Biological Sciences, School of Life Sciences, Ulsan National Institute of Science and Technology (UNIST), Ulsan, 689-798, Korea.
authors.forename: Sebyung
authors.initials: S
authors.lastname: Kang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005978:Glutathione / Q000737:chemistry; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D007700:Kinetics; D051379:Mice; D058525:Quartz Crystal Microbalance Techniques; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D006735:Horseradish Peroxidase; D005982:Glutathione Transferase; D005978:Glutathione