scholarly journals Comparison of MALDI-TOF-MS and RP-HPLC as Rapid Screening Methods for Wheat Lines With Altered Gliadin Compositions

2020 ◽  
Vol 11 ◽  
Author(s):  
You-Ran Jang ◽  
Kyoungwon Cho ◽  
Sewon Kim ◽  
Jae-Ryeong Sim ◽  
Su-Bin Lee ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Mutation Breeding ◽  
Rapid Screening ◽  
Food Allergies ◽  
Screening Methods ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Wheat Lines ◽  
Tof Ms

The wheat gliadins are a complex group of flour proteins that can trigger celiac disease and serious food allergies. As a result, mutation breeding and biotechnology approaches are being used to develop new wheat lines with reduced immunogenic potential. Key to these efforts is the development of rapid, high-throughput methods that can be used as a first step in selecting lines with altered gliadin contents. In this paper, we optimized matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and reversed-phase high-performance liquid chromatography (RP-HPLC) methods for the separation of gliadins from Triticum aestivum cv. Chinese Spring (CS). We evaluated the quality of the resulting profiles using the complete set of gliadin gene sequences recently obtained from this cultivar as well as a set of aneuploid lines in CS. The gliadins were resolved into 13 peaks by MALDI-TOF-MS. α- or γ-gliadins that contain abundant celiac disease epitopes and are likely targets for efforts to reduce the immunogenicity of flour were found in several peaks. However, other peaks contained multiple α- and γ-gliadins, including one peak with as many as 12 different gliadins. In comparison, separation of proteins by RP-HPLC yielded 28 gliadin peaks, including 13 peaks containing α-gliadins and eight peaks containing γ-gliadins. While the separation of α- and γ-gliadins gliadins achieved by RP-HPLC was better than that achieved by MALDI-TOF-MS, it was not possible to link peaks with individual protein sequences. Both MALDI-TOF-MS and RP-HPLC provided adequate separation of ω-gliadins. While MALDI-TOF-MS is faster and could prove useful in studies that target specific gliadins, RP-HPLC is an effective method that can be applied more broadly to detect changes in gliadin composition.

scholarly journals Detection of Ovarian Cancer Using Samples Sourced from the Vaginal Microenvironment

2019 ◽  
Author(s):  
Melissa M. Galey ◽  
Alexandria N. Young ◽  
Valentina Z. Petukhova ◽  
Mingxun Wang ◽  
Jian Wang ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Disease Progression ◽  
Xenograft Model ◽  
Screening Methods ◽  
Maldi Tof Ms ◽  
Analytical Technique ◽  
Maldi Tof ◽  
Specificity And Sensitivity ◽  
Biological Mixtures ◽  
Tof Ms

AbstractMass spectrometry (MS) offers high levels of specificity and sensitivity in clinical applications, and we have previously been able to demonstrate that matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS is capable of distinguishing two-component biological mixtures at low limits of detection. Ovarian cancer is notoriously difficult to detect due to the lack of any screening methods for early detection. By sampling a local microenvironment, such as the vaginal fluids, a MS based method is presented that was capable of monitoring disease progression from vaginally collected, local samples from tumor bearing mice. A murine xenograft model of high grade serous ovarian carcinoma (HGSOC) was used for this study and vaginal lavages were obtained from mice on a weekly basis throughout disease progression and subjected to our MALDI-TOF MS workflow followed by statistical analyses. Proteins in the 4-20 kDa region of the mass spectrum could consistently be measured to yield a fingerprint that correlated with disease progression over time. These fingerprints were found to be statistically stable across all mice with the protein fingerprint converging towards the end point of the study. MALDI-TOF MS serves as a unique analytical technique for measuring a sampled vaginal microenvironment in a specific and sensitive manner for the detection of HGSOC in a murine model.

scholarly journals Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat

Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4347
Author(s):  
You-Ran Jang ◽  
Kyoungwon Cho ◽  
Se Won Kim ◽  
Susan B. Altenbach ◽  
Sun-Hyung Lim ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Throughput ◽  
Glutenin Subunits ◽  
Maldi Tof Ms ◽  
Molecular Weights ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Group 2 ◽  
Tof Ms

Because high-molecular-weight glutenin subunits (HMW-GS) are important contributors to wheat end-use quality, there is a need for high-throughput identification of HMW-GS in wheat genetic resources and breeding lines. We developed an optimized method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to distinguish individual HMW-GS by considering the effects of the alkylating reagent in protein extraction, solvent components, dissolving volume, and matrix II components. Using the optimized method, 18 of 22 HMW-GS were successfully identified in standard wheat cultivars by differences in molecular weights or by their associations with other tightly linked subunits. Interestingly, 1Bx7 subunits were divided into 1Bx7 group 1 and 1Bx7 group 2 proteins with molecular weights of about 82,400 and 83,000 Da, respectively. Cultivars containing the 1Bx7 group 2 proteins were distinguished from those containing 1Bx7OE using well-known DNA markers. HMW-GS 1Ax2* and 1Bx6 and 1By8 and 1By8*, which are difficult to distinguish due to very similar molecular weights, were easily identified using RP-HPLC. To validate the method, HMW-GS from 38 Korean wheat varieties previously evaluated by SDS-PAGE combined with RP-HPLC were analyzed by MALDI-TOF-MS. The optimized MALDI-TOF-MS method will be a rapid, high-throughput tool for selecting lines containing desirable HMW-GS for breeding efforts.

Subunit Characteristics of Pig Pancreas Ferritin Revealed by MALDI-TOF MS and RP-HPLC

2008 ◽  
Vol 24 (5) ◽  
pp. 550-556 ◽  
Author(s):  
L HUANG ◽  
Z LIN ◽  
Q LIN ◽  
L LUO ◽  
H HUANG
Keyword(s):  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Tof Ms

Rapid screening of marine bacterial symbionts using MALDI-TOF MS

2020 ◽  
Vol 202 (8) ◽  
pp. 2329-2336
Author(s):  
Livia M. R. Vidal ◽  
Tainá M. Venas ◽  
Aline R. P. Gonçalves ◽  
Hannah K. Mattsson ◽  
Raphael V. P. Silva ◽  
...  
Keyword(s):  
Rapid Screening ◽  
Bacterial Symbionts ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

scholarly journals Direct Identification of Urinary Tract Pathogens from Urine Samples, Combining Urine Screening Methods and Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

2016 ◽  
Vol 54 (4) ◽  
pp. 988-993 ◽  
Author(s):  
Melania Íñigo ◽  
Andreu Coello ◽  
Gema Fernández-Rivas ◽  
Belén Rivaya ◽  
Jessica Hidalgo ◽  
...  
Keyword(s):  
Laser Desorption ◽  
Urine Samples ◽  
Screening Methods ◽  
Gram Negative Bacteria ◽  
Laser Desorption Ionization ◽  
Ionization Time ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Flight Mass Spectrometry ◽  
Tof Ms

Early diagnosis of urinary tract infections (UTIs) is essential to avoid inadequate or unnecessary empirical antibiotic therapy. Microbiological confirmation takes 24 to 48 h. The use of screening methods, such as cytometry and automated microscopic analysis of urine sediment, allows the rapid prediction of negative samples. In addition, matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) is a widely established technique in clinical microbiology laboratories used to identify microorganisms. We evaluated the ability of MALDI-TOF MS to identify microorganisms from direct urine samples and the predictive value of automated analyzers for the identification of microorganisms in urine by MALDI-TOF MS. A total of 451 urine samples from patients with suspected UTIs were first analyzed using the Sysmex UF-1000iflow cytometer, an automatic sediment analyzer with microscopy (SediMax), culture, and then processed by MALDI-TOF MS with a simple triple-centrifuged procedure to obtain a pellet that was washed and centrifuged and finally applied directly to the MALDI-TOF MS plate. The organisms in 336 samples were correctly identified, mainly those with Gram-negative bacteria (86.10%). No microorganisms were misidentified, and noCandidaspp. were correctly identified. Regarding the data from autoanalyzers, the best bacteriuria cutoffs were 1,000 and 200 U/μl for UF-1000iand SediMax, respectively. It was concluded that the combination of a urine screening method and MALDI-TOF MS provided a reliable identification from urine samples, especially in those containing Gram-negative bacteria.

895: Proteomic Analysis of Sera from Bladder Cancer Patients with MALDI-TOF-MS

2007 ◽  
Vol 177 (4S) ◽  
pp. 297-297
Author(s):  
Kristina Schwamborn ◽  
Rene Krieg ◽  
Ruth Knüchel-Clarke ◽  
Joachim Grosse ◽  
Gerhard Jakse
Keyword(s):  
Bladder Cancer ◽  
Cancer Patients ◽  
Proteomic Analysis ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms
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