title: Comparative surface plasmon resonance and enzyme-linked immunosorbent assay characterisation of a monoclonal antibody with N-acyl homoserine lactones.
abstract: This study shows the detection of (N-acyl) homoserine lactones (AHLs or HSL) with monoclonal antibodies via a surface plasmon resonance (SPR)-based immunosensor in comparison to conventional microtiter plate-based enzyme-linked immunosorbent assay (ELISA). An HSL derivative, named HSL2 (Table 1), was attached to bovine serum albumin (BSA) and the conjugate (HSL2-BSA-r2) was either covalently immobilised on the SPR sensor chip surface via free amino groups or via adsorption on the ELISA polystyrene plate surface. With a newly developed rat monoclonal antibody (mAb HSL1/2 2C10), AHLs were detected sensitively in a competitive format with SPR and ELISA. Well comparable experiments between SPR and ELISA could be obtained in buffers. Moreover, the SPR sensor surface with the immobilised conjugate HSL2-BSA-r2 could be regenerated at least 340 times (regeneration cycles) without loss of activity. The measurement time per cycle was approximately 15 min. The competitive detection format for SPR and ELISA allowed the detection in the μgL(-1) range.
authors.affiliation: Institute of Ecological Chemistry, Helmholtz Center Munich - German Research Center for Environmental Health (GmbH), Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Wöllner
authors.forename: X
authors.initials: X
authors.lastname: Chen
authors.forename: E
authors.initials: E
authors.lastname: Kremmer
authors.forename: P M
authors.initials: PM
authors.lastname: Krämer
mesh_terms: D054742:Acyl-Butyrolactones / Q000032:analysis* / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D001419:Bacteria / Q000378:metabolism; D001667:Binding, Competitive; D015374:Biosensing Techniques / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006241:Haptens / Q000737:chemistry; D053038:Quorum Sensing; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D054742:Acyl-Butyrolactones; D000911:Antibodies, Monoclonal; D006241:Haptens; D012710:Serum Albumin, Bovine