Gregory Crescenzo - Academia.edu (original) (raw)

Papers by Gregory Crescenzo

Journal of Materials Chemistry B, 2015

Please note that technical editing may introduce minor changes to the text and/or graphics, which... more Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal's standard Terms & Conditions and the Ethical guidelines still apply. In no event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains.

Methods in molecular biology, 2024

IFAC Proceedings Volumes, 2010

The emergence of surface plasmon resonance-based optical biosensors has facilitated the identific... more The emergence of surface plasmon resonance-based optical biosensors has facilitated the identification of kinetic parameters for various macromolecular interactions (interactions between an analyte and a ligand). To determine these parameters, normally, the ligand is immobilized on the biosensor surface and experiments with arbitrarily chosen periods of analyte and buffer injections, and varying analyte concentrations are performed. The periods of injection and the analyte concentrations could be optimized to reduce experimental time. On the other hand, there are some ligands that can interact with more than one analyte, and the current practice is to test their interactions individually. In this work, it is shown that by injecting multiple analytes at the same time, it is possible to reduce the experimental time needed for kinetic parameter identification.

Frontiers in Bioengineering and Biotechnology, 2016

Coiled-coil interactions: a versatile bioaffinity system for the oriented immobilization and tuna... more Coiled-coil interactions: a versatile bioaffinity system for the oriented immobilization and tunable release of biomolecules from biomaterials. Frontiers in Bioengineering and Biotechnology, p. 1-2.

Journal of Biomedical Materials Research Part A, 2015

In the context of porous bone void filler for oral bone reconstruction, peptides that suppress mi... more In the context of porous bone void filler for oral bone reconstruction, peptides that suppress microbial growth and promote osteoblast function could be used to enhance the performance of a porous bone void filler. We tested the hypothesis that P15-CSP, a novel fusion peptide containing collagen-mimetic osteogenic peptide P15, and competence-stimulating peptide (CSP), a cationic antimicrobial peptide, has emerging properties not shared by P15 or CSP alone. Peptide-coated surfaces were tested for antimicrobial activity toward Streptoccocus mutans, and their ability to promote human mesenchymal stem cell (MSC) attachment, spreading, metabolism, and osteogenesis. In the osteogenesis assay, peptides were coated on tissue culture plastic and on thin films generated by plasma-enhanced chemical vapor deposition to have hydrophilic or hydrophobic character (water contact angles 63°, 42°, and 92°, respectively). S. mutans planktonic growth was specifically inhibited by CSP, whereas biofilm formation was inhibited by P15-CSP. MSC adhesion and actin stress fiber formation was strongly enhanced by CSP, P15-CSP, and fibronectin coatings and modestly enhanced by P15 versus uncoated surfaces. Metabolic assays revealed that CSP was slightly cytotoxic to MSCs. MSCs developed alkaline phosphatase activity on all surfaces, with or without peptide coatings, and consistently deposited the most biomineralized matrix on hydrophilic surfaces coated with P15-CSP. Hydrophobic thin films completely suppressed MSC biomineralization, consistent with previous findings of suppressed osteogenesis on hydrophobic bioplastics. Collective data in this study provide new evidence that P15-CSP has unique dual capacity to suppress biofilm formation, and to enhance osteogenic activity as a coating on hydrophilic surfaces. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2015.

Chemical Science, 2015

We present a novel, aqueous thiol-based conjugation strategy that constitutes an alternative to t... more We present a novel, aqueous thiol-based conjugation strategy that constitutes an alternative to the oxime-click pathway for generating a reactive end-group on chitosan (CS), which could also be applicable to other polymers.

Journal of molecular recognition : JMR, 2012

Surface plasmon resonance-based biosensors are now acknowledged as robust and reliable instrument... more Surface plasmon resonance-based biosensors are now acknowledged as robust and reliable instruments to determine the kinetic parameters related to the interactions between biomolecules. These kinetic parameters are used in screening campaigns: there is a considerable interest in reducing the experimental time, thus improving the throughput of the surface plasmon resonance assays. Kinetic parameters are typically obtained by analyzing data from several injections of a given analyte at different concentrations over a surface where its binding partner has been immobilized. It has been already proven that an iterative optimization approach aiming at determining optimal analyte injections to be performed online can significantly reduce the experimentation time devoted to kinetic parameter determination, without any detrimental effect on their standard errors. In this study, we explore the potential of this iterative optimization approach to further reduce experiment duration by combining ...

J. Mater. Chem. B, 2013

Bioengineering approaches have been intensively applied to create small diameter vascular grafts ... more Bioengineering approaches have been intensively applied to create small diameter vascular grafts using artificial materials. However, a fully successful, high performing and anti-thrombogenic structure has not been achieved yet. In this study, we present the first step of a process aiming at biofunctionalizing previously designed compliant polyethylene terephthalate (PET) scaffolds (Moreno et al., 2011). The main challenge of such a surface modification is to prevent the bulk polymer from any damage, so that it preserves the mechanical properties that the structures have been designed for. In that endeavor, an aminated long-chain polymer (polyvinylamine, PVAm) was used as an aminolysis reagent to get amine (-NH2) moieties only on the very surface of PET. Different reaction conditions were assayed, leading to a large range of amino group densities associated with slight variations of the planar tensile properties. These results were in stark contrast with those generated with a common small diamine substrate (ethylenediamine, EtDA), as the latter yielded a strong degradation of the mechanical properties for comparable amine densities. Tubular mechanical assays were then carried out on PVAm-functionalized PET scaffolds. The latter showed a compliance match with arteries under the chosen reaction conditions, as initially observed for pristine PET tubular scaffolds.

Methods in Molecular Biology, 2014

ACS Applied Materials & Interfaces, 2014

The characteristics of the gate electrode have significant effects on the behavior of organic ele... more The characteristics of the gate electrode have significant effects on the behavior of organic electrochemical transistors (OECTs), which are intensively investigated for applications in the booming field of organic bioelectronics. In this work, high specific surface area activated carbon (AC) was used as gate electrode material in OECTs based on the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) doped with poly(styrenesulfonate) (PSS). We found that the high specific capacitance of the AC gate electrodes leads to high drain-source current modulation in OECTs, while their intrinsic quasi-reference characteristics make unnecessary the presence of an additional reference electrode to monitor the OECT channel potential.

Biomacromolecules, 2014

This study highlights the advantages of chondroitin sulfate (CS) as a sublayer combining selectiv... more This study highlights the advantages of chondroitin sulfate (CS) as a sublayer combining selective low-fouling properties, low-platelet adhesion and pro-adhesive properties on endothelial cells, making CS promising for vascular graft applications. These properties were evaluated by comparing CS with well-known low-fouling coatings such as poly(ethylene glycol) (PEG) and carboxymethylated dextran (CMD), which were covalently grafted on primary amine-rich plasma polymerized (LP) films. Protein adsorption studies by quartz crystal microbalance with dissipation monitoring (QCM-D) and fluorescence measurements showed that CS is as effective as PEG in reducing fibrinogen adsorption (~90% reduction). CS also largely reduced adsorption of bovine serum albumin (BSA) as well as fetal bovine serum (FBS) but to a lower extent than PEG and CMD surfaces (72% vs 85% for BSA and 66% vs 89% for FBS). Whole blood perfusion assays indicated that, while LP surfaces were highly reactive with platelets, PEG, CMD, and CS grafted surfaces drastically decreased platelet adhesion and activation to levels significantly lower than polyethylene terephthalate (PET) surfaces. Finally, while human umbilical vein endothelial cell (HUVEC) adhesion and growth were found to be very limited on PEG and CMD, they were significantly increased on CS compared to that on bare PET and reached similar values as those for tissue culture polystyrene positive controls. Interestingly, HUVEC retention during perfusion with blood was found to be excellent on CS but poor on PET. Overall, our results suggest that the CS surface has the advantage of promoting HUVEC growth and resistance to flow-induced shear stress while preventing fibrinogen and platelet attachment. Such a nonthrombogenic but endothelial-cell adhesive surface is thus promising to limit vascular graft occlusion.

Sensors and Actuators B: Chemical, 2008

We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that... more We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that enables to combine ultra-high phase sensitivity with good signal-to-noise background. The proposed approach uses spatial modulation of s-polarized component by birefringent elements and the extraction of phase-polarization information by Fourier-transform methods. This scheme was tested for monitoring the interactions between an antibody and its biological partner. Our experimental data, collected by amplitude-sensitive and phase-sensitive polarimetry demonstrate that the latter scheme provides at least one order of magnitude improvement in terms of detection limit.

Molecular and Cellular Biology, 2004

The import of proteins containing the peroxisomal targeting signal 1 (PTS1) into the Leishmania g... more The import of proteins containing the peroxisomal targeting signal 1 (PTS1) into the Leishmania glycosome is dependent on the docking of the PTS1-loaded LdPEX5 cytosolic receptor with LdPEX14 on the glycosome surface. Here we show that, in the absence of PTS1, LdPEX5 is a tetramer that is stabilized by two distinct interaction domains; the first is a coiled-coil motif encompassing residues 277 to 310, whereas the second domain is localized to residues 1 to 202. By using microcalorimetry, surface plasmon resonance, and size exclusion chromatography techniques, we show that PTS1 peptide binding to LdPEX5 tetramers promotes their dissociation into dimeric structures, which are stabilized by a coiled-coil interaction. Moreover, we demonstrated that the resulting LdPEX5-PTS1 complex is remarkably stable and exhibits extremely slow dissociation kinetics. However, binding of LdPEX14 to LdPEX5 modulates the LdPEX5-PTS1 affinity as it decreases the thermodynamic dissociation constant for thi...

Microbiology, 2005

InSalmonella enterica, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory s... more InSalmonella enterica, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg2+. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoPHis) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoPHisare compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoPHiswas mostly monomeric, whereas PhoPHis∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoPHisexhibited different spectral properties. The interaction between PhoP or PhoPHisand th...

Journal of Molecular Recognition, 2008

The emergence of surface plasmon resonance-based optical biosensors has facilitated the identific... more The emergence of surface plasmon resonance-based optical biosensors has facilitated the identification of kinetic parameters for various macromolecular interactions. Normally, these parameters are determined from experiments with arbitrarily chosen periods of macromolecule and buffer injections, and varying macromolecule concentrations. Since the choice of these variables is arbitrary, such experiments may not provide the required confidence in identified kinetic parameters expressed in terms of standard errors. In this work, an iterative optimization approach is used to determine the above-mentioned variables so as to reduce the experimentation time, while treating the required standard errors as constraints. It is shown using multiple experimental and simulated data that the desired confidence can be reached with much shorter experiments than those generally performed by biosensor users.

Journal of Molecular Recognition, 2014

In order to improve the throughput of surface plasmon resonance-based biosensors, an on-line iter... more In order to improve the throughput of surface plasmon resonance-based biosensors, an on-line iterative optimization algorithm has been presented aiming at reducing experimental time and material consumption without any loss of confidence on kinetic parameters [De Crescenzo (2008) J. Mol Recognit., 21, 256-66.]. This algorithm was based on a simple Langmuirian model to compute the confidence and predict optimal injections. However, this kinetic model is not suitable for all interactions, as it does not include mass transfer limitation that may occur for fast interaction kinetics. If a simple model was to be used when this phenomenon influenced the interactions, kinetic parameters would be biased. On the other hand, we show in this paper that data analysis with a kinetic model including a mass transfer limitation step would lead to longer experiments and poorer confidence if the interactions were simple. So, in this manuscript, we present an on-line model discrimination and optimization approach to increase the throughput of surface plasmon resonance biosensors.

Journal of Materials Chemistry B, 2015

Please note that technical editing may introduce minor changes to the text and/or graphics, which... more Please note that technical editing may introduce minor changes to the text and/or graphics, which may alter content. The journal's standard Terms & Conditions and the Ethical guidelines still apply. In no event shall the Royal Society of Chemistry be held responsible for any errors or omissions in this Accepted Manuscript or any consequences arising from the use of any information it contains.

Methods in molecular biology, 2024

IFAC Proceedings Volumes, 2010

The emergence of surface plasmon resonance-based optical biosensors has facilitated the identific... more The emergence of surface plasmon resonance-based optical biosensors has facilitated the identification of kinetic parameters for various macromolecular interactions (interactions between an analyte and a ligand). To determine these parameters, normally, the ligand is immobilized on the biosensor surface and experiments with arbitrarily chosen periods of analyte and buffer injections, and varying analyte concentrations are performed. The periods of injection and the analyte concentrations could be optimized to reduce experimental time. On the other hand, there are some ligands that can interact with more than one analyte, and the current practice is to test their interactions individually. In this work, it is shown that by injecting multiple analytes at the same time, it is possible to reduce the experimental time needed for kinetic parameter identification.

Frontiers in Bioengineering and Biotechnology, 2016

Coiled-coil interactions: a versatile bioaffinity system for the oriented immobilization and tuna... more Coiled-coil interactions: a versatile bioaffinity system for the oriented immobilization and tunable release of biomolecules from biomaterials. Frontiers in Bioengineering and Biotechnology, p. 1-2.

Journal of Biomedical Materials Research Part A, 2015

In the context of porous bone void filler for oral bone reconstruction, peptides that suppress mi... more In the context of porous bone void filler for oral bone reconstruction, peptides that suppress microbial growth and promote osteoblast function could be used to enhance the performance of a porous bone void filler. We tested the hypothesis that P15-CSP, a novel fusion peptide containing collagen-mimetic osteogenic peptide P15, and competence-stimulating peptide (CSP), a cationic antimicrobial peptide, has emerging properties not shared by P15 or CSP alone. Peptide-coated surfaces were tested for antimicrobial activity toward Streptoccocus mutans, and their ability to promote human mesenchymal stem cell (MSC) attachment, spreading, metabolism, and osteogenesis. In the osteogenesis assay, peptides were coated on tissue culture plastic and on thin films generated by plasma-enhanced chemical vapor deposition to have hydrophilic or hydrophobic character (water contact angles 63°, 42°, and 92°, respectively). S. mutans planktonic growth was specifically inhibited by CSP, whereas biofilm formation was inhibited by P15-CSP. MSC adhesion and actin stress fiber formation was strongly enhanced by CSP, P15-CSP, and fibronectin coatings and modestly enhanced by P15 versus uncoated surfaces. Metabolic assays revealed that CSP was slightly cytotoxic to MSCs. MSCs developed alkaline phosphatase activity on all surfaces, with or without peptide coatings, and consistently deposited the most biomineralized matrix on hydrophilic surfaces coated with P15-CSP. Hydrophobic thin films completely suppressed MSC biomineralization, consistent with previous findings of suppressed osteogenesis on hydrophobic bioplastics. Collective data in this study provide new evidence that P15-CSP has unique dual capacity to suppress biofilm formation, and to enhance osteogenic activity as a coating on hydrophilic surfaces. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2015.

Chemical Science, 2015

We present a novel, aqueous thiol-based conjugation strategy that constitutes an alternative to t... more We present a novel, aqueous thiol-based conjugation strategy that constitutes an alternative to the oxime-click pathway for generating a reactive end-group on chitosan (CS), which could also be applicable to other polymers.

Journal of molecular recognition : JMR, 2012

Surface plasmon resonance-based biosensors are now acknowledged as robust and reliable instrument... more Surface plasmon resonance-based biosensors are now acknowledged as robust and reliable instruments to determine the kinetic parameters related to the interactions between biomolecules. These kinetic parameters are used in screening campaigns: there is a considerable interest in reducing the experimental time, thus improving the throughput of the surface plasmon resonance assays. Kinetic parameters are typically obtained by analyzing data from several injections of a given analyte at different concentrations over a surface where its binding partner has been immobilized. It has been already proven that an iterative optimization approach aiming at determining optimal analyte injections to be performed online can significantly reduce the experimentation time devoted to kinetic parameter determination, without any detrimental effect on their standard errors. In this study, we explore the potential of this iterative optimization approach to further reduce experiment duration by combining ...

J. Mater. Chem. B, 2013

Bioengineering approaches have been intensively applied to create small diameter vascular grafts ... more Bioengineering approaches have been intensively applied to create small diameter vascular grafts using artificial materials. However, a fully successful, high performing and anti-thrombogenic structure has not been achieved yet. In this study, we present the first step of a process aiming at biofunctionalizing previously designed compliant polyethylene terephthalate (PET) scaffolds (Moreno et al., 2011). The main challenge of such a surface modification is to prevent the bulk polymer from any damage, so that it preserves the mechanical properties that the structures have been designed for. In that endeavor, an aminated long-chain polymer (polyvinylamine, PVAm) was used as an aminolysis reagent to get amine (-NH2) moieties only on the very surface of PET. Different reaction conditions were assayed, leading to a large range of amino group densities associated with slight variations of the planar tensile properties. These results were in stark contrast with those generated with a common small diamine substrate (ethylenediamine, EtDA), as the latter yielded a strong degradation of the mechanical properties for comparable amine densities. Tubular mechanical assays were then carried out on PVAm-functionalized PET scaffolds. The latter showed a compliance match with arteries under the chosen reaction conditions, as initially observed for pristine PET tubular scaffolds.

Methods in Molecular Biology, 2014

ACS Applied Materials & Interfaces, 2014

The characteristics of the gate electrode have significant effects on the behavior of organic ele... more The characteristics of the gate electrode have significant effects on the behavior of organic electrochemical transistors (OECTs), which are intensively investigated for applications in the booming field of organic bioelectronics. In this work, high specific surface area activated carbon (AC) was used as gate electrode material in OECTs based on the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) doped with poly(styrenesulfonate) (PSS). We found that the high specific capacitance of the AC gate electrodes leads to high drain-source current modulation in OECTs, while their intrinsic quasi-reference characteristics make unnecessary the presence of an additional reference electrode to monitor the OECT channel potential.

Biomacromolecules, 2014

This study highlights the advantages of chondroitin sulfate (CS) as a sublayer combining selectiv... more This study highlights the advantages of chondroitin sulfate (CS) as a sublayer combining selective low-fouling properties, low-platelet adhesion and pro-adhesive properties on endothelial cells, making CS promising for vascular graft applications. These properties were evaluated by comparing CS with well-known low-fouling coatings such as poly(ethylene glycol) (PEG) and carboxymethylated dextran (CMD), which were covalently grafted on primary amine-rich plasma polymerized (LP) films. Protein adsorption studies by quartz crystal microbalance with dissipation monitoring (QCM-D) and fluorescence measurements showed that CS is as effective as PEG in reducing fibrinogen adsorption (~90% reduction). CS also largely reduced adsorption of bovine serum albumin (BSA) as well as fetal bovine serum (FBS) but to a lower extent than PEG and CMD surfaces (72% vs 85% for BSA and 66% vs 89% for FBS). Whole blood perfusion assays indicated that, while LP surfaces were highly reactive with platelets, PEG, CMD, and CS grafted surfaces drastically decreased platelet adhesion and activation to levels significantly lower than polyethylene terephthalate (PET) surfaces. Finally, while human umbilical vein endothelial cell (HUVEC) adhesion and growth were found to be very limited on PEG and CMD, they were significantly increased on CS compared to that on bare PET and reached similar values as those for tissue culture polystyrene positive controls. Interestingly, HUVEC retention during perfusion with blood was found to be excellent on CS but poor on PET. Overall, our results suggest that the CS surface has the advantage of promoting HUVEC growth and resistance to flow-induced shear stress while preventing fibrinogen and platelet attachment. Such a nonthrombogenic but endothelial-cell adhesive surface is thus promising to limit vascular graft occlusion.

Sensors and Actuators B: Chemical, 2008

We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that... more We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that enables to combine ultra-high phase sensitivity with good signal-to-noise background. The proposed approach uses spatial modulation of s-polarized component by birefringent elements and the extraction of phase-polarization information by Fourier-transform methods. This scheme was tested for monitoring the interactions between an antibody and its biological partner. Our experimental data, collected by amplitude-sensitive and phase-sensitive polarimetry demonstrate that the latter scheme provides at least one order of magnitude improvement in terms of detection limit.

Molecular and Cellular Biology, 2004

The import of proteins containing the peroxisomal targeting signal 1 (PTS1) into the Leishmania g... more The import of proteins containing the peroxisomal targeting signal 1 (PTS1) into the Leishmania glycosome is dependent on the docking of the PTS1-loaded LdPEX5 cytosolic receptor with LdPEX14 on the glycosome surface. Here we show that, in the absence of PTS1, LdPEX5 is a tetramer that is stabilized by two distinct interaction domains; the first is a coiled-coil motif encompassing residues 277 to 310, whereas the second domain is localized to residues 1 to 202. By using microcalorimetry, surface plasmon resonance, and size exclusion chromatography techniques, we show that PTS1 peptide binding to LdPEX5 tetramers promotes their dissociation into dimeric structures, which are stabilized by a coiled-coil interaction. Moreover, we demonstrated that the resulting LdPEX5-PTS1 complex is remarkably stable and exhibits extremely slow dissociation kinetics. However, binding of LdPEX14 to LdPEX5 modulates the LdPEX5-PTS1 affinity as it decreases the thermodynamic dissociation constant for thi...

Microbiology, 2005

InSalmonella enterica, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory s... more InSalmonella enterica, PhoP is the response regulator of the PhoP/PhoQ two-component regulatory system that controls the expression of various virulence factors in response to external Mg2+. Previous studies have shown that phosphorylation of a PhoP variant with a C-terminal His tag (PhoPHis) enhances dimerization and binding to target DNA. Here, the effect of phosphorylation on the oligomerization and DNA binding properties of both wild-type PhoP (PhoP) and PhoPHisare compared. Gel filtration chromatography showed that PhoP exists as a mixture of monomer and dimer regardless of its phosphorylation state. In contrast, unphosphorylated PhoPHiswas mostly monomeric, whereas PhoPHis∼P existed as a mixture of monomer and dimer. By monitoring the tryptophan fluorescence of the proteins and the fluorescence of the probe 1-anilinonaphthalene-8-sulfonic acid bound to them, it was found that PhoP and PhoPHisexhibited different spectral properties. The interaction between PhoP or PhoPHisand th...

Journal of Molecular Recognition, 2008

The emergence of surface plasmon resonance-based optical biosensors has facilitated the identific... more The emergence of surface plasmon resonance-based optical biosensors has facilitated the identification of kinetic parameters for various macromolecular interactions. Normally, these parameters are determined from experiments with arbitrarily chosen periods of macromolecule and buffer injections, and varying macromolecule concentrations. Since the choice of these variables is arbitrary, such experiments may not provide the required confidence in identified kinetic parameters expressed in terms of standard errors. In this work, an iterative optimization approach is used to determine the above-mentioned variables so as to reduce the experimentation time, while treating the required standard errors as constraints. It is shown using multiple experimental and simulated data that the desired confidence can be reached with much shorter experiments than those generally performed by biosensor users.

Journal of Molecular Recognition, 2014

In order to improve the throughput of surface plasmon resonance-based biosensors, an on-line iter... more In order to improve the throughput of surface plasmon resonance-based biosensors, an on-line iterative optimization algorithm has been presented aiming at reducing experimental time and material consumption without any loss of confidence on kinetic parameters [De Crescenzo (2008) J. Mol Recognit., 21, 256-66.]. This algorithm was based on a simple Langmuirian model to compute the confidence and predict optimal injections. However, this kinetic model is not suitable for all interactions, as it does not include mass transfer limitation that may occur for fast interaction kinetics. If a simple model was to be used when this phenomenon influenced the interactions, kinetic parameters would be biased. On the other hand, we show in this paper that data analysis with a kinetic model including a mass transfer limitation step would lead to longer experiments and poorer confidence if the interactions were simple. So, in this manuscript, we present an on-line model discrimination and optimization approach to increase the throughput of surface plasmon resonance biosensors.