PMID- 37268197
OWN - NLM
STAT- MEDLINE
DCOM- 20230621
LR  - 20230621
IS  - 1096-0309 (Electronic)
IS  - 0003-2697 (Linking)
VI  - 674
DP  - 2023 Aug 1
TI  - Step-by-step full factorial design to optimize a quantitative sandwich ELISA.
PG  - 115195
LID - S0003-2697(23)00160-4 [pii]
LID - 10.1016/j.ab.2023.115195 [doi]
AB  - In this work, a quantitative sandwich ELISA was optimized, through a full 
      factorial design of experiments (DOE) in successive steps of a preliminary 
      protocol obtained by the method of one factor at a time (OFAT). The specificity 
      of the optimized ELISA, the lower limit of quantification, the quantification 
      range and the analytical sensitivity of the antigen quantification curve were 
      evaluated, in comparison with the curve obtained from the preliminary protocol. 
      The full factorial DOE was linked to a simple statistical processing, which 
      facilitates the interpretation of the results in those laboratories where there 
      is no trained statistician. The step-by-step optimization of the ELISA and the 
      successive incorporation into the protocol of the best combination of factors and 
      levels, allowed obtaining a specific immunoassay, with an analytical sensitivity 
      20 times greater and with a lower limit of antigen quantification that decreased 
      from 156.25 at 9.766 ng/mL. As far as we know, there are no reports of 
      optimization of an ELISA following the step-by-step scheme used in this work. The 
      optimized ELISA will be used for the quantification of the TT-P0 protein, the 
      active principle of a vaccine candidate against sea lice.
CI  - Copyright (c) 2023 Elsevier Inc. All rights reserved.
FAU - Hernandez, Carlos A
AU  - Hernandez CA
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Perez-Bernal, Maylin
AU  - Perez-Bernal M
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba. Electronic address: 
      maylin.perez@cigb.edu.cu.
FAU - Abreu, Daymi
AU  - Abreu D
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Valdivia, Onel
AU  - Valdivia O
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Delgado, Magali
AU  - Delgado M
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Dorta, Dayami
AU  - Dorta D
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Dominguez, Andy G
AU  - Dominguez AG
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Perez, Enrique R
AU  - Perez ER
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
FAU - Sanchez-Rios, Jose M
AU  - Sanchez-Rios JM
AD  - Center for Genetic Engineering and Biotechnology of Sancti Spiritus, 
      Circunvalante Norte, Olivos III, Sancti Spiritus, Cuba.
LA  - eng
PT  - Journal Article
DEP - 20230601
PL  - United States
TA  - Anal Biochem
JT  - Analytical biochemistry
JID - 0370535
RN  - 0 (Antigens)
SB  - IM
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Immunoassay
MH  - *Antigens
OTO - NOTNLM
OT  - ELISA
OT  - Effect size
OT  - Full factorial design
OT  - Optimization
COIS- Declaration of competing interest The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2023/06/03 11:42
MHDA- 2023/06/21 06:42
CRDT- 2023/06/02 19:24
PHST- 2023/03/28 00:00 [received]
PHST- 2023/05/11 00:00 [revised]
PHST- 2023/05/22 00:00 [accepted]
PHST- 2023/06/21 06:42 [medline]
PHST- 2023/06/03 11:42 [pubmed]
PHST- 2023/06/02 19:24 [entrez]
AID - S0003-2697(23)00160-4 [pii]
AID - 10.1016/j.ab.2023.115195 [doi]
PST - ppublish
SO  - Anal Biochem. 2023 Aug 1;674:115195. doi: 10.1016/j.ab.2023.115195. Epub 2023 Jun 
      1.