PMID- 15183767
OWN - NLM
STAT- MEDLINE
DCOM- 20050125
LR  - 20131121
IS  - 0003-2697 (Print)
IS  - 0003-2697 (Linking)
VI  - 330
IP  - 1
DP  - 2004 Jul 1
TI  - Comparative analyses of a small molecule/enzyme interaction by multiple users of 
      Biacore technology.
PG  - 98-113
AB  - To gauge the experimental variability associated with Biacore analysis, 36 
      different investigators analyzed a small molecule/enzyme interaction under 
      similar conditions. Acetazolamide (222 g/mol) binding to carbonic anhydrase II 
      (CAII; 30000 Da) was chosen as a model system. Both reagents were stable and 
      their interaction posed a challenge to measure because of the low molecular 
      weight of the analyte and the fast association rate constant. Each investigator 
      created three different density surfaces of CAII and analyzed an identical 
      dilution series of acetazolamide (ranging from 4.1 to 1000 nM). The greatest 
      variability in the results was observed during the enzyme immobilization step 
      since each investigator provided their own surface activating reagents. 
      Variability in the quality of the acetazolamide binding responses was likely a 
      product of how well the investigators' instruments had been maintained. To 
      determine the reaction kinetics, the responses from the different density 
      surfaces were fit globally to a 1:1 interaction model that included a term for 
      mass transport. The averaged association and dissociation rate constants were 
      3.1+/-1.6 x 10(6)M(-1)s(-1) and 6.7+/-2.5 x 10(-2)s(-1), respectively, which 
      corresponded to an average equilibrium dissociation constant (K(D) of 2.6+/-1.4 x 
      10(-8)M. The results provide a benchmark of variability in interpreting binding 
      constants from the biosensor and highlight keys areas that should be considered 
      when analyzing small molecule interactions.
FAU - Cannon, Michelle J
AU  - Cannon MJ
AD  - Center for Biomolecular Interaction Analysis, University of Utah, Salt Lake City, 
      UT 84132, USA.
FAU - Papalia, Giuseppe A
AU  - Papalia GA
FAU - Navratilova, Iva
AU  - Navratilova I
FAU - Fisher, Robert J
AU  - Fisher RJ
FAU - Roberts, Lindsey R
AU  - Roberts LR
FAU - Worthy, Karen M
AU  - Worthy KM
FAU - Stephen, Andrew G
AU  - Stephen AG
FAU - Marchesini, Gerardo R
AU  - Marchesini GR
FAU - Collins, Edward J
AU  - Collins EJ
FAU - Casper, Dave
AU  - Casper D
FAU - Qiu, Huawei
AU  - Qiu H
FAU - Satpaev, Daulet
AU  - Satpaev D
FAU - Liparoto, Stefano F
AU  - Liparoto SF
FAU - Rice, Dax A
AU  - Rice DA
FAU - Gorshkova, Inna I
AU  - Gorshkova II
FAU - Darling, Ryan J
AU  - Darling RJ
FAU - Bennett, Donald B
AU  - Bennett DB
FAU - Sekar, Michael
AU  - Sekar M
FAU - Hommema, Eric
AU  - Hommema E
FAU - Liang, Amy M
AU  - Liang AM
FAU - Day, Eric S
AU  - Day ES
FAU - Inman, Jean
AU  - Inman J
FAU - Karlicek, Shannon M
AU  - Karlicek SM
FAU - Ullrich, Stephen J
AU  - Ullrich SJ
FAU - Hodges, Dianne
AU  - Hodges D
FAU - Chu, Teresa
AU  - Chu T
FAU - Sullivan, Eric
AU  - Sullivan E
FAU - Simpson, Jack
AU  - Simpson J
FAU - Rafique, Ashique
AU  - Rafique A
FAU - Luginbuhl, Beatrice
AU  - Luginbuhl B
FAU - Westin, Susanne Nyholm
AU  - Westin SN
FAU - Bynum, Magdalena
AU  - Bynum M
FAU - Cachia, Paul
AU  - Cachia P
FAU - Li, Yue-Jin
AU  - Li YJ
FAU - Kao, Daniel
AU  - Kao D
FAU - Neurauter, Amy
AU  - Neurauter A
FAU - Wong, Melanie
AU  - Wong M
FAU - Swanson, Michael
AU  - Swanson M
FAU - Myszka, David G
AU  - Myszka DG
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - EC 4.2.1.- (Carbonic Anhydrase II)
RN  - O3FX965V0I (Acetazolamide)
SB  - IM
MH  - Acetazolamide/*chemistry/metabolism
MH  - Carbonic Anhydrase II/*chemistry/metabolism
MH  - Kinetics
MH  - Observer Variation
MH  - Protein Binding
MH  - Research Personnel
MH  - *Surface Plasmon Resonance/instrumentation/standards
EDAT- 2004/06/09 05:00
MHDA- 2005/01/26 09:00
CRDT- 2004/06/09 05:00
PHST- 2003/12/04 00:00 [received]
PHST- 2004/06/09 05:00 [pubmed]
PHST- 2005/01/26 09:00 [medline]
PHST- 2004/06/09 05:00 [entrez]
AID - S0003269704001782 [pii]
AID - 10.1016/j.ab.2004.02.027 [doi]
PST - ppublish
SO  - Anal Biochem. 2004 Jul 1;330(1):98-113. doi: 10.1016/j.ab.2004.02.027.