TY - JOUR
T1 - Detection of Brucella abortus by a platform functionalized with protein A and specific antibodies IgG
AU - Baltierra-Uribe, Shantal Lizbeth
AU - Chanona-Pérez, José Jorge
AU - Méndez-Méndez, Juan Vicente
AU - Perea-Flores, María de Jesús
AU - Sánchez-Chávez, Anahí Carolina
AU - García-Pérez, Blanca Estela
AU - Moreno-Lafont, Martha Cecilia
AU - López-Santiago, Rubén
N1 - Publisher Copyright:
© 2019 Wiley Periodicals, Inc.
PY - 2019/5
Y1 - 2019/5
N2 - 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.
AB - 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.
KW - antibody orientation
KW - atomic force microscopy
KW - brucellosis
KW - immunosensors
KW - protein A
UR - http://www.scopus.com/inward/record.url?scp=85060030433&partnerID=8YFLogxK
U2 - 10.1002/jemt.23206
DO - 10.1002/jemt.23206
M3 - Artículo
C2 - 30637865
SN - 1059-910X
VL - 82
SP - 586
EP - 595
JO - Microscopy Research and Technique
JF - Microscopy Research and Technique
IS - 5
ER -