PMID- 30802413
OWN - NLM
STAT- MEDLINE
DCOM- 20200413
LR  - 20231213
IS  - 2472-5560 (Electronic)
IS  - 2472-5552 (Linking)
VI  - 24
IP  - 5
DP  - 2019 Jun
TI  - A General Guide for the Optimization of Enzyme Assay Conditions Using the Design 
      of Experiments Approach.
PG  - 587-596
LID - 10.1177/2472555219830084 [doi]
AB  - Many factors must be considered during the optimization of an enzyme assay. These 
      include the choice of buffer and its composition, the type of enzyme and its 
      concentration, as well as the type of substrate and concentrations, the reaction 
      conditions, and the appropriate assay technology. The process of an enzyme assay 
      optimization, in our experience, can take more than 12 weeks using the 
      traditional one-factor-at-a-time approach. In contrast, the design of experiments 
      (DoE) approaches have the potential to speed up the assay optimization process 
      and provide a more detailed evaluation of tested variables. However, not all 
      researchers are aware of DoE approaches or believe that it is easy to employ a 
      DoE approach for the optimization of an assay. In order to facilitate enzyme 
      assay developers to use DoE methodologies, we present in detail the steps 
      required to identify in less than 3 days (1) the factors that significantly 
      affect the activity of an enzyme and (2) the optimal assay conditions using a 
      fractional factorial approach and response surface methodology. This is 
      exemplified with the optimization of assay conditions for the human rhinovirus-3C 
      protease, and the methodology used could be employed as a basic guide for the 
      speedy identification of the optimum assay conditions for any enzyme.
FAU - Onyeogaziri, Favour Chinyere
AU  - Onyeogaziri FC
AD  - 1 Department of Life and Health Sciences, School of Sciences and Engineering, 
      University of Nicosia, Nicosia, Cyprus.
FAU - Papaneophytou, Christos
AU  - Papaneophytou C
AUID- ORCID: 0000-0002-4045-8839
AD  - 1 Department of Life and Health Sciences, School of Sciences and Engineering, 
      University of Nicosia, Nicosia, Cyprus.
LA  - eng
PT  - Journal Article
DEP - 20190225
PL  - United States
TA  - SLAS Discov
JT  - SLAS discovery : advancing life sciences R & D
JID - 101697563
RN  - 0 (Viral Proteins)
RN  - EC 3.4.22.- (Cysteine Endopeptidases)
RN  - EC 3.4.22.28 (3C Viral Proteases)
SB  - IM
MH  - 3C Viral Proteases
MH  - Biological Assay/*methods
MH  - Cysteine Endopeptidases/*chemistry
MH  - Enzyme Assays/*methods
MH  - Humans
MH  - Research Design
MH  - Substrate Specificity
MH  - Viral Proteins/*chemistry
OTO - NOTNLM
OT  - assay optimization
OT  - design of experiments
OT  - enzyme activity
OT  - fractional factorial
OT  - response surface methodology
EDAT- 2019/02/26 06:00
MHDA- 2020/04/14 06:00
CRDT- 2019/02/26 06:00
PHST- 2019/02/26 06:00 [pubmed]
PHST- 2020/04/14 06:00 [medline]
PHST- 2019/02/26 06:00 [entrez]
AID - S2472-5552(22)06779-X [pii]
AID - 10.1177/2472555219830084 [doi]
PST - ppublish
SO  - SLAS Discov. 2019 Jun;24(5):587-596. doi: 10.1177/2472555219830084. Epub 2019 Feb 
      25.