Author/Authors :
Gupta، Garima نويسنده Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, INDIA , , Kumar، Ashu نويسنده Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, INDIA , , Boopathi، Mannan نويسنده , , Thavaselvam، Duraipandian نويسنده Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, INDIA , , Singh ، Beer نويسنده Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, INDIA , , Vijayaraghavan، Rajagopalan نويسنده Defence Research and Development Establishment, Jhansi Road, Gwalior-474002, INDIA ,
Abstract :
A rapid and simple method for the real time monitoring of Brucella abortus
CSP-31 (B. abortus CSP-31) by using Surface Plasmon resonance (SPR) is first time
reported. SPR and electrochemical impedance spectroscopy (EIS) were used to
characterize the B. abortus CSP-31 interaction on carboxymethyldextran modified gold
disc with antibody. The EIS data revealed a decrease in impedance after interaction of
antigen with antibody due to the effective binding. The present SPR biosensor is able to
detect 0.05 pM concentration of B. abortus CSP-31 in phosphate buffered saline in less
than 10 minute with linearity from 2.0 to 16.0 pM. The equilibrium constant (KD) and
maximum binding capacity of analyte (Bmax) values for B. abortus CSP-31 were
calculated to be 7.6 pM and 235.94 m?, respectively. This KD value classifies the antibody
as a high affinity one towards the antigen. Moreover, thermodynamic parameter change in
Gibb’s free energy was calculated and found to be -63.34 kJ/mol. Interference study was
also performed with B. abortus cell envelope (CE) antigen and a negative angle change
was observed with exemplifying specificity for the antigen-antibody interaction of B.
abortus CSP-31.
