PMID- 34342257
OWN - NLM
STAT- MEDLINE
DCOM- 20220818
LR  - 20220921
IS  - 1875-5402 (Electronic)
IS  - 1386-2073 (Linking)
VI  - 25
IP  - 9
DP  - 2022
TI  - Systematic Optimisation of Microtiter Plate Lectin Assay to Improve Sialic Acid 
      Linkage Detection.
PG  - 1507-1517
LID - 10.2174/1386207324666210802122538 [doi]
AB  - AIMS: We aimed to develop a high-throughput lectin assay with minimized 
      background signals to investigate the interactions of lectins and sialic acid 
      glycans, focusing on Prostate- Specific Antigen (PSA). BACKGROUND: High 
      background signals resulting from nonspecific binding are a significant concern 
      for microtiter plate-based Enzyme-Linked Lectin Sorbent Assays (ELISAs), as they 
      can mask specific binding signals and cause false-positive results. METHODS: In 
      this study, we constructed an ELISA based on different washing step parameters, 
      including the number of washing cycles, NaCl and Tween-20 concentrations, and the 
      type of blocking agent and evaluated the effects on both specific and nonspecific 
      binding signals. Furthermore, we performed a PSA binding assay using the 
      optimized ELISA. RESULTS: The optimal washing parameters based on the highest 
      specific binding signal proposed four cycles of washing steps using a washing 
      buffer containing a high salt concentration (0.5 M NaCl) and mild detergent 
      (0.05% Tween-20). The utilization of the optimized washing parameters in this 
      assay was shown to be sufficient to obtain the optimal binding signals without 
      the use of any blocking agent. Binding assays performed using the optimized ELISA 
      revealed that the glycan of the PSA sample used in this study mainly consists of 
      terminal alpha2,6-linked sialic acid, as strongly recognized by Sambucus nigra 
      agglutinin (SNA) with a KD value of 12.38 nM. CONCLUSION: The ELISA reported in 
      this study provides a simple yet sensitive assay for sialic acid linkage 
      recognition.
CI  - Copyright(c) Bentham Science Publishers; For any queries, please email at 
      epub@benthamscience.net.
FAU - Khairol Mokhtar, Nur Hanina Izzati
AU  - Khairol Mokhtar NHI
AD  - School of Biological Science and Biotechnology, Faculty of Science and 
      Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor 43600, Malaysia.
FAU - Hussin, Ainulkhir
AU  - Hussin A
AD  - Department of Pathology, Queen Elizabeth Hospital, Ministry of Health, Kota 
      Kinabalu, Sabah, Malaysia.
FAU - Hamid, Aidil Abdul
AU  - Hamid AA
AD  - School of Biological Science and Biotechnology, Faculty of Science and 
      Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor 43600, Malaysia.
FAU - Zainal Ariffin, Shahrul Hisham
AU  - Zainal Ariffin SH
AD  - School of Biological Science and Biotechnology, Faculty of Science and 
      Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor 43600, Malaysia.
FAU - Shahidan, Muhammad Ashraf
AU  - Shahidan MA
AD  - School of Biological Science and Biotechnology, Faculty of Science and 
      Technology, Universiti Kebangsaan Malaysia, Bangi, Selangor 43600, Malaysia.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United Arab Emirates
TA  - Comb Chem High Throughput Screen
JT  - Combinatorial chemistry & high throughput screening
JID - 9810948
RN  - 0 (Lectins)
RN  - 0 (Polysaccharides)
RN  - 0 (Polysorbates)
RN  - 451W47IQ8X (Sodium Chloride)
RN  - EC 3.4.21.77 (Prostate-Specific Antigen)
RN  - GZP2782OP0 (N-Acetylneuraminic Acid)
SB  - IM
MH  - Humans
MH  - *Lectins/metabolism
MH  - Male
MH  - *N-Acetylneuraminic Acid
MH  - Polysaccharides
MH  - Polysorbates
MH  - Prostate-Specific Antigen/metabolism
MH  - Sodium Chloride
OTO - NOTNLM
OT  - Sialic acid
OT  - enzyme-linked lectin sorbent assay
OT  - glycans
OT  - high-throughput
OT  - lectins
OT  - prostate-specific antigen
EDAT- 2021/08/04 06:00
MHDA- 2022/08/19 06:00
CRDT- 2021/08/03 08:38
PHST- 2020/12/30 00:00 [received]
PHST- 2021/06/02 00:00 [revised]
PHST- 2021/06/04 00:00 [accepted]
PHST- 2021/08/04 06:00 [pubmed]
PHST- 2022/08/19 06:00 [medline]
PHST- 2021/08/03 08:38 [entrez]
AID - CCHTS-EPUB-116982 [pii]
AID - 10.2174/1386207324666210802122538 [doi]
PST - ppublish
SO  - Comb Chem High Throughput Screen. 2022;25(9):1507-1517. doi: 
      10.2174/1386207324666210802122538.