Development of Novel Monoclonal Antibodies to Wheat Alpha-Amylases Associated with Grain Quality Problems That Are Increasing with Climate Change
Abstract
:1. Introduction
2. Results
2.1. Expression of Analysis of Wheat (Triticum astevium; TA) α-Amylase (AMY) Genes
2.2. Monoclonal Antibody Development
2.3. Validation of TaAMY Antibodies for α-Amylase Detection
2.4. Validation of TaAMY Antibody Specificity
2.5. Evaluating the Use TaAMY Antibodies in Pairs
3. Discussion
3.1. Overview
3.2. Expression Analysis of Wheat α-Amylase Genes
3.3. Potential for an α-Amylase Immunoassay for Wheat
4. Materials and Methods
4.1. Plant Material and Growth Conditions
4.2. Expression Analysis of α-Amylase Genes in Grain Development
4.3. Cloning of α-Amylase Genes and Recombinant Protein Expression
4.4. Statistical Analysis
4.5. Peptide Monoclonal Antibody Development
4.6. Induction of α-Amylase Enzymes from Aleurone Tissue
4.7. Monoclonal Antibody Validation and Peptide Sequencing
4.8. Alpha-Amylase Immunoprecipitation Assays
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Hauvermale, A.L.; Matzke, C.; Bohaliga, G.; Pumphrey, M.O.; Steber, C.M.; McCubbin, A.G. Development of Novel Monoclonal Antibodies to Wheat Alpha-Amylases Associated with Grain Quality Problems That Are Increasing with Climate Change. Plants 2023, 12, 3798. https://doi.org/10.3390/plants12223798
Hauvermale AL, Matzke C, Bohaliga G, Pumphrey MO, Steber CM, McCubbin AG. Development of Novel Monoclonal Antibodies to Wheat Alpha-Amylases Associated with Grain Quality Problems That Are Increasing with Climate Change. Plants. 2023; 12(22):3798. https://doi.org/10.3390/plants12223798
Chicago/Turabian StyleHauvermale, Amber L., Courtney Matzke, Gamila Bohaliga, Mike O. Pumphrey, Camille M. Steber, and Andrew G. McCubbin. 2023. "Development of Novel Monoclonal Antibodies to Wheat Alpha-Amylases Associated with Grain Quality Problems That Are Increasing with Climate Change" Plants 12, no. 22: 3798. https://doi.org/10.3390/plants12223798