Detection of Brucella abortus by a platform functionalized with protein A and specific antibodies IgG

Oriented immobilization of antibodies on a sensor surface is critical for enhancing both the antigen‐binding capacity and the sensitivity of immunosensors. In this study, we describe a strategy to adsorb immunoglobulin G (IgG) anti‐Brucella antibodies onto a silicon surface, oriented by protein A ob...

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Bibliographic Details
Published in:Microscopy research and technique 2019-05, Vol.82 (5), p.586-595
Main Authors: Baltierra‐Uribe, Shantal Lizbeth, Chanona‐Pérez, José Jorge, Méndez‐Méndez, Juan Vicente, Perea‐Flores, María de Jesús, Sánchez‐Chávez, Anahí Carolina, García‐Pérez, Blanca Estela, Moreno‐Lafont, Martha Cecilia, López‐Santiago, Rubén
Format: Article
Language:English
Subjects:
Antibodies
antibody orientation
Antigens
Atomic beam spectroscopy
Atomic force microscopy
Biosensors
Brucella
Brucella abortus
brucellosis
Confocal microscopy
Immobilization
Immunoglobulin G
Immunoglobulins
Immunosensors
Microscopes
Microscopy
Organic chemistry
Photoelectron spectroscopy
Photoelectrons
Protein A
Proteins
Silicon
Spectroscopy
Spectrum analysis
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Summary:Oriented immobilization of antibodies on a sensor surface is critical for enhancing both the antigen‐binding capacity and the sensitivity of immunosensors. In this study, we describe a strategy to adsorb immunoglobulin G (IgG) anti‐Brucella antibodies onto a silicon surface, oriented by protein A obtained from Staphylococcus aureus (SpA). X‐ray photoelectron spectroscopy and atomic force microscopy were used to characterize topographically, morphologically, and chemical changes of the sensor functionalization. The activity of the biosensor was assessed by confocal microscopy, scanning electronic microscopy, and bacteria capture assays (BCA). According to the BCA, the efficiency of Brucella abortus detection with the SpA‐IgG anti Brucella biosensor was three‐fold higher than that of the random orientated IgG anti Brucella biosensor. The limit of detection was 1 × 106 CFU/ml. These data show that the orientation of antibodies immobilization is crucial to developing immunosensors for bacterial antigen detection as Brucella spp and improve its sensibility level. Functionalization with protein A increases Brucella detection by an antibody‐coated surface. Functionalized silicon surface for Brucella detection was characterized by atomic force microscopy, X‐ray photoelectron spectroscopy and confocal microscopy.
ISSN:1059-910X
1097-0029
DOI:10.1002/jemt.23206