PMID- 20734499
OWN - NLM
STAT- MEDLINE
DCOM- 20101122
LR  - 20191111
IS  - 1520-5207 (Electronic)
IS  - 1520-5207 (Linking)
VI  - 114
IP  - 32
DP  - 2010 Aug 19
TI  - Investigation of an allergen adsorption on amine- and acid-terminated thiol 
      layers: influence on their affinity to specific antibodies.
PG  - 10612-9
AB  - This work describes the controlled immobilization of a recognized allergen, 
      beta-lactoglobulin, onto gold transducers with the aim of optimizing the 
      elaboration of a biosensor directed against allergen-produced antibodies. This 
      protein was immobilized on both amine- and acid-terminated thiol self-assembled 
      monolayers, and the influence on its affinity to a specific IgG was investigated. 
      For amine-terminated layers, the beta-lactoglobulin was immobilized via its 
      surface acid functions implying an activation step with 
      1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride/ester of 
      N-hydroxysuccinimide (EDC-NHS). Conversely, the grafting on acid-terminated layer 
      takes advantage of the accessible amine groups that react with the activated 
      acidalkylthiols. The resulting layers of beta-lactoglobulin were then submitted 
      to various concentrations of rabbit serum containing beta-lactoglobulin specific 
      rabbit immunoglobulin (rIgG), and the antigen/antibody affinity was evaluated 
      using modulated polarization-infrared absorption spectroscopy (PM-IRRAS) and 
      Fourier transform surface plasmon resonance (FT-SPR). Even though for similar 
      concentration, the amount of adsorbed beta-lactoglobulin was identical on both 
      surfaces, atomic force microscopy (AFM) images showed a better dispersion for 
      amine-terminated layers. Moreover, the affinity to specific IgG, estimated under 
      static conditions by PM-IRRAS and under dynamic conditions by SPR, was different. 
      Grafting beta-lactoglobulin via its acid groups gave an affinity constant 3 times 
      higher than its immobilization via its amine groups despite the fact that the 
      amount of accessible recognition sites appeared to be similar for both systems. 
      This work underlines the importance of the involved chemical groups upon protein 
      immobilization on their biological activity and will be essential for the 
      construction of nondirect biosensors for detecting specific immunoglobulin E 
      (IgE) of allergens.
FAU - Thebault, Pascal
AU  - Thebault P
AD  - UPMC Univ. Paris 6, UMR 7197, Laboratoire de Reactivite de Surface, 4 Place 
      Jussieu, F75005 Paris, France.
FAU - Boujday, Souhir
AU  - Boujday S
FAU - Senechal, Helene
AU  - Senechal H
FAU - Pradier, Claire-Marie
AU  - Pradier CM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Phys Chem B
JT  - The journal of physical chemistry. B
JID - 101157530
RN  - 0 (Allergens)
RN  - 0 (Amines)
RN  - 0 (Antibodies)
RN  - 0 (Lactoglobulins)
RN  - 0 (Sulfhydryl Compounds)
RN  - 7440-57-5 (Gold)
SB  - IM
MH  - Adsorption
MH  - *Allergens/chemistry/immunology
MH  - Amines/*chemistry
MH  - Animals
MH  - Antibodies/*immunology
MH  - *Antibody Affinity
MH  - Biosensing Techniques
MH  - Child
MH  - Gold/chemistry
MH  - Humans
MH  - *Lactoglobulins/chemistry/immunology
MH  - Microscopy, Atomic Force
MH  - Rabbits
MH  - Spectrum Analysis/methods
MH  - Sulfhydryl Compounds/*chemistry
EDAT- 2010/08/25 06:00
MHDA- 2010/12/14 06:00
CRDT- 2010/08/25 06:00
PHST- 2010/08/25 06:00 [entrez]
PHST- 2010/08/25 06:00 [pubmed]
PHST- 2010/12/14 06:00 [medline]
AID - 10.1021/jp103164b [doi]
PST - ppublish
SO  - J Phys Chem B. 2010 Aug 19;114(32):10612-9. doi: 10.1021/jp103164b.