PMID- 12054440
OWN - NLM
STAT- MEDLINE
DCOM- 20021217
LR  - 20111117
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 305
IP  - 2
DP  - 2002 Jun 15
TI  - A fluorescent-photochrome method for the quantitative characterization of solid 
      phase antibody orientation.
PG  - 121-34
AB  - A fluorescent-photochrome method of quantifying the orientation and surface 
      density of solid phase antibodies is described. The method is based on 
      measurements of quenching and rates of cis-trans photoisomerization and 
      photodestruction of a stilbene-labeled hapten by a quencher in solution. These 
      experimental parameters enable a quantitative description of the order of binding 
      sites of antibodies immobilized on a surface and can be used to characterize the 
      microviscosity and steric hindrance in the vicinity of the binding site. 
      Furthermore, a theoretical method for the determination of the depth of immersion 
      of the fluorescent label in a two-phase system was developed. The model exploits 
      the concept of dynamic interactions and is based on the empirical dependence of 
      parameters of static exchange interactions on distances between exchangeable 
      centers. In the present work, anti-dinitrophenyl (DNP) antibodies and 
      stilbene-labeled DNP were used to investigate three different protein 
      immobilization methods: physical adsorption, covalent binding, and the 
      Langmuir-Blodgett technique.
CI  - Copyright 2002 Elsevier Science (USA).
FAU - Ahluwalia, Arti
AU  - Ahluwalia A
AD  - Interdepartmental Research Center "E. Piaggio", via Diotisalvi 2, Pisa, 56126, 
      Italy.
FAU - De Rossi, Danilo
AU  - De Rossi D
FAU - Giusto, Giuseppe
AU  - Giusto G
FAU - Chen, Oren
AU  - Chen O
FAU - Papper, Vladislav
AU  - Papper V
FAU - Likhtenshtein, Gertz I
AU  - Likhtenshtein GI
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Antibodies)
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Iodides)
RN  - 0 (Silanes)
RN  - 0 (Stilbenes)
RN  - 11121-56-5 (Phytochrome)
SB  - IM
MH  - Adsorption
MH  - Antibodies/*analysis/chemistry
MH  - Binding Sites, Antibody
MH  - Computational Biology
MH  - Dinitrobenzenes/immunology
MH  - Fluorescent Dyes/*metabolism
MH  - Iodides/metabolism
MH  - Kinetics
MH  - Models, Molecular
MH  - Phytochrome/*metabolism
MH  - Silanes/metabolism
MH  - Stilbenes/immunology
EDAT- 2002/06/11 10:00
MHDA- 2002/12/18 04:00
CRDT- 2002/06/11 10:00
PHST- 2002/06/11 10:00 [pubmed]
PHST- 2002/12/18 04:00 [medline]
PHST- 2002/06/11 10:00 [entrez]
AID - S0003269702956010 [pii]
AID - 10.1006/abio.2002.5601 [doi]
PST - ppublish
SO  - Anal Biochem. 2002 Jun 15;305(2):121-34. doi: 10.1006/abio.2002.5601.