Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance

Background: A method for the selection of suitable molecular recognition element (MRE) for the quantification of human epidermal growth factor (hEGF) using surface plasmon resonance (SPR) is presented. Two types of hEGF antibody, monoclonal and polyclonal, were immobilized on the surface of chip and...

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Autores principales:	Rosti,Ira Amira, Ramanan,Ramakrishnan Nagasundara, Ling,Tau Chuan, Ariff,Arbakariya B
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2013
Materias:	antibody BIAcore biosensors epidermal growth factor validation
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600015
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spelling	oai:scielo:S0717-345820130006000152014-01-16Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonanceRosti,Ira AmiraRamanan,Ramakrishnan NagasundaraLing,Tau ChuanAriff,Arbakariya B antibody BIAcore biosensors epidermal growth factor validation Background: A method for the selection of suitable molecular recognition element (MRE) for the quantification of human epidermal growth factor (hEGF) using surface plasmon resonance (SPR) is presented. Two types of hEGF antibody, monoclonal and polyclonal, were immobilized on the surface of chip and validated for its characteristics and performance in the quantification of hEGF. Validation of this analytical procedure was to demonstrate the stability and suitability of antibody for the quantification of target protein. Results: Specificity, accuracy and precision for all samples were within acceptable limit for both antibodies. The affinity and kinetic constant of antibodies-hEGF binding were evaluated using a 1:1 Langmuir interaction model. The model fitted well to all binding responses simultaneously. Polyclonal antibody (pAb) has better affinity (K D = 7.39e-10 M) than monoclonal antibody (mAb) (K D = 9.54e-9 M). Further evaluation of kinetic constant demonstrated that pAb has faster reaction rate during sample injection, slower dissociation rate during buffer injection and higher level of saturation state than mAb. Besides, pAb has longer shelf life and greater number of cycle run. Conclusions: Thus, pAb was more suitable to be used as a stable MRE for further quantification works from the consideration of kinetic, binding rate and shelf life assessment.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.16 n.6 20132013-11-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600015en10.2225/vol16-issue6-fulltext-11
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	antibody BIAcore biosensors epidermal growth factor validation
spellingShingle	antibody BIAcore biosensors epidermal growth factor validation Rosti,Ira Amira Ramanan,Ramakrishnan Nagasundara Ling,Tau Chuan Ariff,Arbakariya B Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
description	Background: A method for the selection of suitable molecular recognition element (MRE) for the quantification of human epidermal growth factor (hEGF) using surface plasmon resonance (SPR) is presented. Two types of hEGF antibody, monoclonal and polyclonal, were immobilized on the surface of chip and validated for its characteristics and performance in the quantification of hEGF. Validation of this analytical procedure was to demonstrate the stability and suitability of antibody for the quantification of target protein. Results: Specificity, accuracy and precision for all samples were within acceptable limit for both antibodies. The affinity and kinetic constant of antibodies-hEGF binding were evaluated using a 1:1 Langmuir interaction model. The model fitted well to all binding responses simultaneously. Polyclonal antibody (pAb) has better affinity (K D = 7.39e-10 M) than monoclonal antibody (mAb) (K D = 9.54e-9 M). Further evaluation of kinetic constant demonstrated that pAb has faster reaction rate during sample injection, slower dissociation rate during buffer injection and higher level of saturation state than mAb. Besides, pAb has longer shelf life and greater number of cycle run. Conclusions: Thus, pAb was more suitable to be used as a stable MRE for further quantification works from the consideration of kinetic, binding rate and shelf life assessment.
author	Rosti,Ira Amira Ramanan,Ramakrishnan Nagasundara Ling,Tau Chuan Ariff,Arbakariya B
author_facet	Rosti,Ira Amira Ramanan,Ramakrishnan Nagasundara Ling,Tau Chuan Ariff,Arbakariya B
author_sort	Rosti,Ira Amira
title	Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
title_short	Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
title_full	Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
title_fullStr	Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
title_full_unstemmed	Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
title_sort	assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2013
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000600015
work_keys_str_mv	AT rostiiraamira assessmentofmolecularrecognitionelementforthequantificationofhumanepidermalgrowthfactorusingsurfaceplasmonresonance AT ramananramakrishnannagasundara assessmentofmolecularrecognitionelementforthequantificationofhumanepidermalgrowthfactorusingsurfaceplasmonresonance AT lingtauchuan assessmentofmolecularrecognitionelementforthequantificationofhumanepidermalgrowthfactorusingsurfaceplasmonresonance AT ariffarbakariyab assessmentofmolecularrecognitionelementforthequantificationofhumanepidermalgrowthfactorusingsurfaceplasmonresonance
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Assessment of molecular recognition element for the quantification of human epidermal growth factor using surface plasmon resonance

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