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Applications of Analytical Chemistry in Biological and Medical Sciences
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Applications of Analytical Chemistry in Biological and Medical Sciences

A special issue of Applied Sciences (ISSN 2076-3417). This special issue belongs to the section "Applied Biosciences and Bioengineering".

Deadline for manuscript submissions: closed (20 August 2022) | Viewed by 23466

Special Issue Editor

Prof. Carlo Zambonin

E-Mail Website
Guest Editor
Department Chemistry, Universuty of Bari Aldo Moro, Via Orabona 4, I-70125 Bari, Italy
Interests: liquid chromatography; gas chromatography; mass spectrometry; maldi
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

It is a pleasure to participate as a Guest Editor in the design of a Special Issue of "Applied Sciences" entitled “Applications of Analytical Chemistry in Biological and Medical Sciences”. Analytical chemistry plays a key role both in the fields of biology and medicine. The analysis of molecules in biological samples, including blood, urine, and tissue, is crucial to understand biological phenomena. The development and validation of innovative analytical methods play critical roles in the development of new drugs and the monitoring of drugs after “in vivo” administration. The discovery of novel tumor markers as well as the monitoring of the levels of existing ones are essential for diagnostic and prognostic evaluations in various kinds of malignant diseases. The use of advanced analytical techniques in the field of proteomics has provided efficient methods for generating sequences, leading to the maximization of the potential of connecting genetic and protein sequences. Based on the above considerations, the aim of the present Issue is to collect original research and review articles reporting a wide range of analytical applications performed in biomedical matrices. Therefore, authors are invited to submit relevant contributions until the submission deadline.

Prof. Carlo Zambonin
Guest Editor

Manuscript Submission Information

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Keywords

  • analytical chemistry
  • biological samples
  • mass spectrometry
  • chromatography
  • sample preparation
  • extraction techniques
  • analytical methods
  • proteomic
  • lipidomic
  • metabolomic

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Published Papers (7 papers)

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Research

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12 pages, 2198 KiB  
Article
Celector®: An Innovative Technology for Quality Control of Living Cells
by Silvia Zia, Barbara Roda, Alessia Maggio, Pasquale Marrazzo, Valeria Pizzuti, Francesco Alviano, Laura Bonsi, Valentina Marassi, Andrea Zattoni and Pierluigi Reschiglian
Appl. Sci. 2022, 12(19), 9967; https://doi.org/10.3390/app12199967 - 4 Oct 2022
Cited by 4 | Viewed by 1601
Abstract
Among the in vitro and ex vivo models used to study human cancer biology, cancer cell lines are widely utilized. The standardization of a correct tumor model including the stage of in vitro testing would allow for the development of new high-efficiency drug [...] Read more.
Among the in vitro and ex vivo models used to study human cancer biology, cancer cell lines are widely utilized. The standardization of a correct tumor model including the stage of in vitro testing would allow for the development of new high-efficiency drug systems. The poor correlation between preclinical in vitro and in vivo data and clinical trials is still an open issue, hence the need for new systems for the quality control (QC) of these cell products. In this work, we present a new technology, Celector®, capable of the label-free analysis and separation of cells based on their physical characteristics with full preservation of their native properties. Two types of cancer cell lines were used: HL60 as cells growing in suspension and SW620 as adherent cells. Cell lines in general show a growth variability depending on the passage and method of culture. Celector® highlights physical differences that can be correlated to cell viability. This work demonstrates the use of Celector® as an analytical platform for the QC of cells used for drug screening, with fundamental improvement of preclinical tests. Cells with a stable doubling time under analysis can be collected and used as standardized systems for high-quality drug monitoring. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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12 pages, 2988 KiB  
Article
Tracking Heme-Protein Interactions in Healthy and Pathological Human Serum in Native Conditions by Miniaturized FFF-Multidetection
by Valentina Marassi, Stefano Giordani, Pierluigi Reschiglian, Barbara Roda and Andrea Zattoni
Appl. Sci. 2022, 12(13), 6762; https://doi.org/10.3390/app12136762 - 4 Jul 2022
Cited by 15 | Viewed by 2638
Abstract
The interaction of heme with blood serum proteins plays an important role in many physiological and pathological processes involving enzyme activity, gene expression and cell proliferation. The mechanisms underlying these interactions are; however, not yet fully understood. New analytical methods able to investigate [...] Read more.
The interaction of heme with blood serum proteins plays an important role in many physiological and pathological processes involving enzyme activity, gene expression and cell proliferation. The mechanisms underlying these interactions are; however, not yet fully understood. New analytical methods able to investigate protein-heme binding in native, biologically representative conditions are thus required. In this work, we present a method based on miniaturized, hollow-fiber flow field-flow fractionation with multiple spectrophotometric and light-scattering detection for size separation of high-abundance serum proteins and selective detection of heme-bound subpopulations. Heme is found to mainly interact with serum albumin, whereas a low amount also binds to other proteins such as IgM. The ability to bind heme in physiological conditions is also investigated for individual serum proteins. IgG is found unable to bind heme at clinically relevant concentrations. The proposed method allows separation, quantitation, and mass/size characterization of serum high-abundance proteins, providing information of heme-protein complex stability and preferred heme-clearing pathways. The same approach could be in perspective extended to the investigation of specific heme-antibody binding, and to further studies involving other molecules of pharmaceutical/clinical interest. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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11 pages, 1917 KiB  
Article
The Effect of Incubation near an Inversely Oriented Square Pyramidal Structure on Adsorption Properties of Horseradish Peroxidase
by Yuri D. Ivanov, Vadim Yu. Tatur, Tatyana O. Pleshakova, Ivan D. Shumov, Andrey F. Kozlov, Anastasia A. Valueva, Irina A. Ivanova, Maria O. Ershova, Nina D. Ivanova, Igor N. Stepanov, Andrei A. Lukyanitsa and Vadim S. Ziborov
Appl. Sci. 2022, 12(8), 4042; https://doi.org/10.3390/app12084042 - 16 Apr 2022
Cited by 3 | Viewed by 1883
Abstract
The incubation of a solution of horseradish peroxidase (HRP) enzyme either below the apex or near the base of an inversely oriented square pyramid (inverted square pyramid; ISP) has been found to influence the enzyme’s aggregation and adsorption properties. The HRP enzyme is [...] Read more.
The incubation of a solution of horseradish peroxidase (HRP) enzyme either below the apex or near the base of an inversely oriented square pyramid (inverted square pyramid; ISP) has been found to influence the enzyme’s aggregation and adsorption properties. The HRP enzyme is used herein as a model object due to its importance in analytical chemistry applications. Atomic force microscopy (AFM) is employed to investigate the HRP’s adsorption on mica substrates at the single-molecule level. Conventional spectrophotometry is used in parallel as a reference method for the determination of the HRP’s enzymatic activity. Using AFM, we reveal a significant change in the adsorption properties of HRP on mica substrates after the incubation of the HRP solutions either above the base or below the apex of the ISP in comparison with the control HRP solution. The same situation is observed after the incubation of the enzyme solution above the center of the ISP’s base. Here, the enzymatic activity of HRP remained unaffected in both cases. Since pyramidal structures of positive and inverted orientation are employed in biosensor devices, it is important to take into account the results obtained herein in the development of highly sensitive biosensor systems, in which pyramidal structures are employed as sensor (such as AFM probes) or construction elements. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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9 pages, 1466 KiB  
Article
Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method
by Seo-jin Park, Byeong ill Lee, Min-Ho Park, Jangmi Choi, Yuri Park, Min-jae Park, Jeong-hyeon Lim, Jiyu Lee, Sangsoo Hwang, Jeongmin Lee and Young G. Shin
Appl. Sci. 2021, 11(20), 9437; https://doi.org/10.3390/app11209437 - 11 Oct 2021
Cited by 2 | Viewed by 3126
Abstract
Trastuzumab emtansine (T-DM1, brand name: Kadcyla®) is the first FDA-approved antibody-drug conjugate (ADC) for metastatic human epidermal growth factor receptor 2 positive (HER2+) breast cancer. It consists of three components: trastuzumab, an anti-HER2 monoclonal antibody, maytansinoid (DM1) as a cytotoxic drug, [...] Read more.
Trastuzumab emtansine (T-DM1, brand name: Kadcyla®) is the first FDA-approved antibody-drug conjugate (ADC) for metastatic human epidermal growth factor receptor 2 positive (HER2+) breast cancer. It consists of three components: trastuzumab, an anti-HER2 monoclonal antibody, maytansinoid (DM1) as a cytotoxic drug, and maleimidomethyl cyclohexane-1-carboxylate (MCC) as a linker. In particular, the MCC linker is known as a non-cleavable linker and has a feature of being conjugated to DM1 by a covalent thioether bond. In this study, we developed an immuno-affinity capture liquid chromatography-mass spectrometric (LC-MS/MS) assay for quantifying the antibody-conjugated drug (acDrug) component of T-DM1. To quantify acDrug, desulfurated DM1 was prepared using a chemical desulfuration pretreatment and quantified as an acDrug. A quadratic regression (weighted 1/concentration), with equation y = ax2 + bx + c, was used to fit the calibration curves over the concentration range of 17.09~1709.44 ng/mL for the acDrug of T-DM1. The quantification run met the in-house acceptance criteria of ±25% accuracy and precision values for the quality control (QC) samples. In conclusion, an immuno-affinity capture LC-MS/MS assay was successfully developed to quantify acDrug of T-DM1 and applied to evaluate in vitro plasma linker stability and preclinical pharmacokinetic (PK) study in rats. This assay could be helpful when applied to other ADCs with the same linker-cytotoxic drug platform. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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11 pages, 10365 KiB  
Article
Determination of VOCs in Surgical Resected Tissues from Colorectal Cancer Patients by Solid Phase Microextraction Coupled to Gas Chromatography–Mass Spectrometry
by Nicoletta De Vietro, Antonella Maria Aresta, Arcangelo Picciariello, Maria Teresa Rotelli and Carlo Zambonin
Appl. Sci. 2021, 11(15), 6910; https://doi.org/10.3390/app11156910 - 27 Jul 2021
Cited by 5 | Viewed by 1824
Abstract
Early diagnosis of colorectal cancer is crucial to increase the survival rates of the patients and breath analysis represents a promising non-invasive tool to obtain information on cancer-related variations on the human volatilome. A solid phase microextraction coupled to gas chromatography–mass spectrometry method [...] Read more.
Early diagnosis of colorectal cancer is crucial to increase the survival rates of the patients and breath analysis represents a promising non-invasive tool to obtain information on cancer-related variations on the human volatilome. A solid phase microextraction coupled to gas chromatography–mass spectrometry method for the determination of seven selected compounds, representative of the volatilome secreted by the colonic mucosa of patients affected by colorectal cancer, including benzaldehyde, benzoic acid, dodecane, ethylbenzene, octanal, tetradecane and toluene, was developed. All the extraction parameters were studied for both headspace and direct immersion sampling and the procedures fully validated. The potential of the approach was demonstrated by the time monitoring of the emission of the selected volatile organic compounds from the surgical resected colon mucosa tissues of colorectal cancer patients. Furthermore, the extraction and identification of thirty-one volatile organic compounds secreted by the same tissues was accomplished. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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Review

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13 pages, 269 KiB  
Review
MALDI-TOF/MS Analysis of Extracellular Vesicles Released by Cancer Cells
by Carlo Zambonin
Appl. Sci. 2022, 12(12), 6149; https://doi.org/10.3390/app12126149 - 16 Jun 2022
Cited by 6 | Viewed by 2208
Abstract
The direct shedding of extracellular vesicles (EVs) from the plasma membrane is a recognized fundamental method for the intercellular transfer of properties in both physiological and pathological conditions. EVs are classified according to origin, biogenesis, size, content, surface markers, and/or functional properties, and [...] Read more.
The direct shedding of extracellular vesicles (EVs) from the plasma membrane is a recognized fundamental method for the intercellular transfer of properties in both physiological and pathological conditions. EVs are classified according to origin, biogenesis, size, content, surface markers, and/or functional properties, and contain various bioactive molecules depending on the physiological state and the type of the cells of origin including lipids, nucleic acids, and proteins. The presence of tumor-derived EVs in body fluids such as blood, ascites, urine, and saliva, together with the important role played in the tumor microenvironment where they intervene at different levels from oncogenesis to metastasis, make EVs a priority target for cancer studies. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can play a leading role in the analysis and characterization of EVs and their load due to its intrinsic advantages such as high throughput, low sample consumption, speed, the cost-effectiveness of the analysis, and the ease of use. This work reviews the main MALDI-TOF applications for the analysis and characterization of extracellular vesicles in the tumor field. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
14 pages, 887 KiB  
Review
MALDI-Based Mass Spectrometry in Clinical Testing: Focus on Bacterial Identification
by Sachio Tsuchida and Tomohiro Nakayama
Appl. Sci. 2022, 12(6), 2814; https://doi.org/10.3390/app12062814 - 9 Mar 2022
Cited by 23 | Viewed by 7413
Abstract
The term “proteome” refers to the total of all proteins expressed in an organism. The term “proteomics” refers to the field of research that includes not only information on the expression levels of individual proteins, but also their higher-order structures, intermolecular interactions, and [...] Read more.
The term “proteome” refers to the total of all proteins expressed in an organism. The term “proteomics” refers to the field of research that includes not only information on the expression levels of individual proteins, but also their higher-order structures, intermolecular interactions, and post-translational modifications. The core technology, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), is available for protein analysis thanks to the work of Koichi Tanaka and John Fenn, who were awarded the Nobel Prize in Chemistry in 2002. The most successful proteome analysis in clinical practice is rapid microbial identification. This method determines the bacterial species by comparing the proteome profile of the bacteria obtained by matrix-assisted laser desorption ionization-time of flight MS (MALDI-TOF MS) with a database. MS is superior in simplicity, speed, and accuracy to classic speciation by staining and phenotyping. In clinical microbiology, MS has had a large impact on the diagnosis and treatment of infectious disease. Early diagnosis and treatment of infectious disease are important, and rapid identification by MALDI-TOF MS has made a major contribution to this field. Full article
(This article belongs to the Special Issue Applications of Analytical Chemistry in Biological and Medical Sciences)
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