Preliminary Studies on Cell-free Fetal DNA Separation and Extraction in Glass Lab-on-a-chip for Capillary Gel Electrophoresis

Background: Beta thalassemia is a common disorder with autosomal recessive inheritance. The most prenatal diagnostic methods are the invasive techniques that have the risk of miscarriage. Now the non-invasive methods will be gradually alternative for these invasive techniques. Objective: The aim of this study is to evaluate and compare the diagnostic value of two non-invasive diagnostic methods for fetal thalassemia using cell free fetal DNA (cff-DNA) and nucleated RBC (NRBC) in one sampling community. Methods: 10 ml of blood was taken in two k3EDTA tube from 32 pregnant women (mean of gestational age = 11 weeks), who themselves and their husbands had minor thalassemia. One tube was used to enrich NRBC and other was used for cff-DNA extraction. NRBCs were isolated by MACS method and immunohistochemistry; the genome of stained cells was amplified by multiple displacement amplification (MDA) procedure. These products were used as template in b-globin segments PCR. cff-DNA was extracted by THP method and 300 bp areas were recovered from the agarose gel as fetus DNA. These DNA were used as template in touch down PCR to amplify b-globin gen. The amplified b-globin segments were sequenced and the results compared with CVS resul. Results: The data showed that sensitivity and specificity of thalassemia diagnosis by NRBC were 100% and 92% respectively and sensitivity and specificity of thalassemia diagnosis by cff-DNA were 100% and 84% respectively. Conclusion: These methods with high sensitivity can be used as screening test but due to their lower specificity than CVS, they cannot be used as diagnostic test.

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Effect of the Monomer Cross-Linker Ratio on the Separation Selectivity of Monoclonal Antibody Subunits in Sodium Dodecyl Sulfate Capillary Gel Electrophoresis

Analytical Chemistry ◽

10.1021/acs.analchem.0c04927 ◽

2021 ◽

Vol 93 (7) ◽

pp. 3535-3541

Author(s):

Csenge Filep ◽

András Guttman

Keyword(s):

Monoclonal Antibody ◽

Sodium Dodecyl Sulfate ◽

Gel Electrophoresis ◽

Sodium Dodecyl ◽

Capillary Gel Electrophoresis ◽

Separation Selectivity ◽

Dodecyl Sulfate ◽

Cross Linker

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Rapid capillary gel electrophoresis analysis of human milk oligosaccharides for food additive manufacturing in-process control

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-020-03119-0 ◽

2021 ◽

Author(s):

Marton Szigeti ◽

Agnes Meszaros-Matwiejuk ◽

Dora Molnar-Gabor ◽

Andras Guttman

Keyword(s):

Additive Manufacturing ◽

Process Control ◽

Human Milk ◽

Gel Electrophoresis ◽

Process Analysis ◽

Food Additive ◽

Human Milk Oligosaccharides ◽

Milk Oligosaccharides ◽

Capillary Gel Electrophoresis

AbstractIndustrial production of human milk oligosaccharides (HMOs) represents a recently growing interest since they serve as key ingredients in baby formulas and are also utilized as dietary supplements for all age groups. Despite their short oligosaccharide chain lengths, HMO analysis is challenging due to extensive positional and linkage variations. Capillary gel electrophoresis primarily separates analyte molecules based on their hydrodynamic volume to charge ratios, thus, offers excellent resolution for most of such otherwise difficult-to-separate isomers. In this work, two commercially available gel compositions were evaluated on the analysis of a mixture of ten synthetic HMOs. The relevant respective separation matrices were then applied to selected analytical in-process control examples. The conventionally used carbohydrate separation matrix was applied for the in-process analysis of bacteria-mediated production of 3-fucosyllactose, lacto-N-tetraose, and lacto-N-neotetraose. The other example showed the suitability of the method for the in vivo in-process control of a shake flask and fermentation approach of 2′-fucosyllactose production. In this latter instance, borate complexation was utilized to efficiently separate the 2′- and 3-fucosylated lactose positional isomers. In all instances, the analysis of the HMOs of interest required only a couple of minutes with high resolution and excellent migration time and peak area reproducibility (average RSD 0.26% and 3.56%, respectively), features representing high importance in food additive manufacturing in-process control. Graphical abstract

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Non-Invasive Prenatal Testing: Current Perspectives and Future Challenges

Genes ◽

10.3390/genes12010015 ◽

2020 ◽

Vol 12 (1) ◽

pp. 15

Author(s):

Luigi Carbone ◽

Federica Cariati ◽

Laura Sarno ◽

Alessandro Conforti ◽

Francesca Bagnulo ◽

...

Keyword(s):

Prenatal Screening ◽

Prenatal Testing ◽

Non Invasive ◽

Fetal Dna ◽

Cell Free Fetal Dna ◽

Common Causes ◽

Future Challenges ◽

Neurodevelopmental Impairment ◽

Fetal Aneuploidies ◽

Made In

Fetal aneuploidies are among the most common causes of miscarriages, perinatal mortality and neurodevelopmental impairment. During the last 70 years, many efforts have been made in order to improve prenatal diagnosis and prenatal screening of these conditions. Recently, the use of cell-free fetal DNA (cff-DNA) testing has been increasingly used in different countries, representing an opportunity for non-invasive prenatal screening of pregnant women. The aim of this narrative review is to describe the state of the art and the main strengths and limitations of this test for prenatal screening of fetal aneuploidies.

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