<p><strong>The team of Professor&nbsp;</strong><a target="_blank" rel="noopener noreferrer" href="https://hocek.group.uochb.cz/en"><strong>Michal Hocek</strong></a><strong> at IOCB Prague explores the limits of the structure and function of DNA and successfully pushes forward known boundaries. An article presenting the results of their latest research has just been published in the major scientific journal </strong><i><strong>Nucleic Acids Research</strong></i><strong>. They prove that even heavily modified double helices of DNA are stable enough to be used for special applications. This discovery might expand the possibilities of utilizing modified DNA, for example, in medicine.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Tomas_Bellon_Scientists_from_IOCB_Prague_are_pushing_the_limits_of_DNA_b5c448f9a9.jpg" alt="ÚOCHB: Scientists from IOCB Prague are pushing the limits of DNA." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Tomas_Bellon_Scientists_from_IOCB_Prague_are_pushing_the_limits_of_DNA_b5c448f9a9.jpg 245w," sizes="100vw" width="1910" height="1000"></p><p>DNA consists of two long negatively charged chains that hold together even though they repel each other. This is made possible by what is known as stacking interactions and base pairing. Michal Hocek and his colleagues advanced the known limits when they modified DNA by adding an extra negative charge to each letter of the genetic code. This doubled the repulsive forces, yet the DNA double helix remained stable. They also found that such modified DNA not only holds together but that it can also be synthesized or replicated and sequenced using the enzyme DNA polymerase.</p><p><i>"Think of DNA as a scaffold on which you can attach various chemical compounds with different functions. These are small molecules, such as side chains of amino acids, which occur naturally only in peptides and proteins. In current medicine, we are able to use these molecules only to a relatively limited extent. The reason is that they are markedly unstable and get decomposed rapidly when inside an organism. One solution to this would be a stable skeleton to which they could be firmly attached. And DNA could be just such a base structure in the future,"</i> says Prof. Michal Hocek.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Tomas_Bellon_Scientists_from_IOCB_Prague_are_pushing_the_limits_of_DNA_Prof_Michal_Hocek_Distinguished_Chair_and_head_of_Bioorganic_and_Medicinal_Chemistry_of_Nucleic_Acids_0a8c924ac4.jpg" alt="ÚOCHB / Tomáš Belloň: Prof. Michal Hocek, Distinguished Chair and head of Bioorganic and Medicinal Chemistry of Nucleic Acids." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Tomas_Bellon_Scientists_from_IOCB_Prague_are_pushing_the_limits_of_DNA_Prof_Michal_Hocek_Distinguished_Chair_and_head_of_Bioorganic_and_Medicinal_Chemistry_of_Nucleic_Acids_0a8c924ac4.jpg 234w," sizes="100vw" width="1500" height="1001"></p><p>The aim of this research carried out at IOCB Prague is to create DNA molecules that will mimic other chemical compounds. This would enable us to leverage the medicinal potential of certain biomolecules that are difficult to retain in the body. Examples of such biomolecules are the already mentioned peptides or proteins.</p><p>The Hocek group has already made significant progress, although research in this direction is at its beginning worldwide. The team has, for example, succeeded at developing a novel modified aptamer, which is a short DNA sequence that can bind to a specific target molecule, most frequently a protein. Aptamers have similar properties to antibodies, but they are much more stable. For this reason, aptamers can potentially replace antibodies employed in medicine. However, the number of approved therapeutic aptamers globally can still be counted on the fingers of one hand.</p><p>The current research exploring the limits of DNA is part of a larger project led by Michal Hocek, for which he has secured a prestigious EXPRO grant from the Czech Science Foundation. This is the second time its results have drawn interest from the journal <i>Nucleic Acids Research</i>. Previously, scientists from IOCB Prague published a method for the enzymatic synthesis of fully artificial DNA. During this synthesis, all nucleotides forming natural DNA are replaced by their modified versions with attached hydrophobic (i.e. water-repelling) molecules.</p>

Scientists from IOCB Prague are pushing the limits of DNA

<p><strong>Researchers from IOCB Prague are furthering the understanding of how medicines work and what it takes to develop their most effective variants. In one current study, they have focused on the disease caused by the protozoan parasite </strong><i><strong>Trichomonas vaginalis</strong></i><strong>, especially because of the recent appearance of strains that are resistant to conventional treatment. In an effort to find a new weak spot of this parasite, the </strong><a target="_blank" rel="noopener noreferrer" href="https://boura.group.uochb.cz/en"><strong>research group</strong></a><strong> led by Dr. Evžen Bouřa has succeeded in preparing a key enzyme complex – the proteasome. This has made it possible to gain knowledge that is indispensable for the development of new effective medicines. The research was carried out in collaboration with colleagues from the University of California San Diego. An article presenting the work has been published in the scientific journal </strong><i><strong>Nature Communications</strong></i><strong>.</strong></p><p>The proteasome is a complex protease that works as a device for recycling old proteins in cells. It is a cylindrical enzyme complex composed of seven subunits arranged in four superposed circles. When old proteins enter this cylinder, they get broken down into small pieces later used by the cell to make new proteins. If it functions incorrectly, old proteins accumulate, leading to the cell’s demise. This is why the blocking of proteasome activity is used, for example, against certain types of malignant tumours.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Tomas_Bellon_Vyzkum_vedcu_z_UOCHB_pomaha_vylepsovat_ucinky_lecivych_latek_Dr_Evzen_Boura_vedouci_vedecke_skupiny_Strukturni_biologie_membran_UOCHB_783ec20050.jpg" alt="IOCB Prague / Tomáš Belloň: Dr. Evžen Bouřa, head of the Structural Membrane Biology group at IOCB Prague." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Tomas_Bellon_Vyzkum_vedcu_z_UOCHB_pomaha_vylepsovat_ucinky_lecivych_latek_Dr_Evzen_Boura_vedouci_vedecke_skupiny_Strukturni_biologie_membran_UOCHB_783ec20050.jpg 234w," sizes="100vw" width="1500" height="1001"></p><div><p>In the same way, it should also be possible to treat the widespread sexually transmitted infection trichomoniasis, which increases the risk of contracting HIV. The aim is therefore to identify substances that block the function of the proteasome inside <i>Trichomonas vaginalis</i> cells. However, the proteasome of this single-cell organism had never been satisfactorily isolated, so the research group from IOCB Prague prepared an artificial proteasome that is identical to the natural one. Curiously, they used insect cells to do so. This part of their research, on which the success of the whole project was hinged, proved the most complicated. That the team has prevailed in describing the previously unknown structure of the parasite’s proteasome is therefore of critical importance.</p><p><i>‘The proteasome looks the same in every animal or protozoan, but the details of this cylindrical structure differ. It is these details that we need to know in order to create a substance that is toxic to the parasite but not harmful to the human body. That is a crucial precondition for drug development. Our greatest success is that we have been able to prepare a recombinant, artificial proteasome that can provide us with the necessary information,’</i> explains Evžen Bouřa.</p><p>The researchers used advanced electron microscopy, allowing them to see almost every single atom. Armed with this capability, they tested the effects of two potential proteasome-inhibiting substances. They saw how they bind to the active site of the proteasome and under what circumstances the proteasome stops working, which is fatal to the cell of the parasite.</p><p>The research was carried out within the <a target="_blank" rel="noopener noreferrer" href="https://nivb.cz/en/">National Institute of Virology and Bacteriology</a> (LX22NPO5103), funded by the <a target="_blank" rel="noopener noreferrer" href="https://www.uochb.cz/en/exceles">EXCELES</a> program.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Vyzkum_vedcu_z_UOCHB_pomaha_vylepsovat_ucinky_lecivych_latek_1_8ff5bf4e71.jpg" alt="IOCB Prague: Dr. Evžen Bouřa, head of the Structural Membrane Biology group at IOCB Prague." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Vyzkum_vedcu_z_UOCHB_pomaha_vylepsovat_ucinky_lecivych_latek_1_8ff5bf4e71.jpg 245w," sizes="100vw" width="1386" height="632"></p><p>&nbsp;</p></div>

Scientists from IOCB Prague help to improve medical drugs

<p><strong>In the week of October 14, the following webinars await you in the field of liquid phase and especially HPLC and LC/MS.</strong></p><blockquote><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinars"><strong>👉 DETAILS AND REGISTRATION IN THE WEBINARS SECTION</strong></a></p><p><strong>👉 Search and filter in the largest global database</strong> with over <strong>3 500 GC, LC, MS and spectroscopy webinars</strong>.</p></blockquote><h5><strong>1. Metrohm: Electrochemical techniques for battery research and testing</strong></h5><ul><li><div><div>Tu, 15.10.2024 10:00 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4337">Registration</a></div></li></ul><p>Theoretical and practical aspects of various electrochemical techniques for battery research and testing.</p><h5><strong>2.&nbsp;Waters Corporation: Leveraging light scattering techniques for enhanced biotherapeutic characterization and development</strong></h5><ul><li><div><div>Tu, 15.10.2024 10:30 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4336">Registration</a></div></li></ul><p>How different light scattering techniques can be used for advanced characterization of biopharmaceutical samples.</p><h5><strong>3.&nbsp;Thermo Fisher Scientific: Solutions for environmental and food contaminant testing</strong></h5><div><ul><li><div>Tu, 15.10.2024 10:30 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4238">Registration</a></div></li></ul></div><p>New and advancing workflows to meet and exceed European regulations.</p><h5><strong>4.&nbsp;Agilent Technologies: Single Quad mass detection for Chromatographers</strong></h5><ul><li><div><div>Tu, 15.10.2024 11:00 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4261">Registration</a></div></li></ul><p>We will look at the development of single quads together and highlight the possibilities offered by today's single quads and how they support the user.</p><h5><strong>5.&nbsp;Agilent Technologies: MassHunter Acquisition Q-TOF Part II – Advance Settings</strong></h5><div><div><ul><li><div>Tu, 15.10.2024 14:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4343">Registration</a></div></li></ul></div><p>Join us for an in-depth exploration of advanced settings in MassHunter Acquisition for Q-TOF systems.</p></div><h5><strong>6.&nbsp;Thermo Fisher Scientific: High-throughput and automation in novel modalities (msAbs, ADCs, AOCs) screening, characterization, and beyond</strong></h5><ul><li><div><div>Tu, 15.10.2024 17:00 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4352">Registration</a></div></li></ul><p>Integrated advanced techniques such as automation and machine learning to achieve unprecedented mass spectrometry-focused sample handling capabilities and rapid turnover rates.</p><h5><strong>7.&nbsp;Phenomenex: Solid Phase Extraction Troubleshooting Tips and Tricks</strong></h5><ul><li><div><div>Tu, 15.10.2024 19:00 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4338">Registration</a></div></li></ul><p>Identify the common challenges associated with solid phase extraction methods.</p><h5><strong>8.&nbsp;Waters Corporation: Streamlined Proteomics with the Power of Andrew+</strong></h5><ul><li><div><div>We, 16.10.2024 11:00 CEST</div></div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4339">Registration</a></div></li></ul><p>How the Andrew+ liquid handling robot has helped streamline their sample preparation workflows for proteomic analysis.</p><h5><strong>9.&nbsp;Agilent Technologies: Advancements in Host Cell Protein Analysis</strong></h5><div><ul><li><div>We, 16.10.2024 11:30 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4357">Registration</a></div></li></ul></div><p>Join us in this unique webinar where experts from Industry, pharmacopeia and Agilent will share insights on latest advancements on HCP analysis.</p><h5><strong>10.&nbsp;Waters Corporation: Free Testosterone and Free T3/T4</strong></h5><div><ul><li><div>We, 16.10.2024 14:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4356">Registration</a></div></li></ul></div><p>Simplified Equilibrium Dialysis and SPE Method for the UPLC-MS/MS Analysis of Free Thyroxine (FT4) and Triiodothyronine (FT3) for Clinical Research.</p><h5><strong>11.&nbsp;Technology Networks/Thermo Fisher Scientific: Streamlining Data for Superior Therapeutic Manufacturing</strong></h5><div><ul><li><div>We, 16.10.2024 17:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4377">Registration</a></div></li></ul></div><p>This webinar explores how to implement digitization to accelerate decision-making, meet regulatory demands and eliminate bottlenecks in biotherapeutic development.</p><h5><strong>12.&nbsp;Waters Corporation: Analyzing mRNA and sgRNA with Advanced Enzymes, Mass Spectrometry, and Informatics</strong></h5><div><ul><li><div>We, 16.10.2024 17:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4355">Registration</a></div></li></ul></div><p>This webinar will feature insights from Quality Assistance a leading European Contract Research Organisation (CRO) offering a range of services including mRNA and CRISPR sgRNA testing.</p><h5><strong>13.&nbsp;Separation Science/Merck: Empowering Tomorrow’s Therapeutics with LC-MS Capillary Chromatography</strong></h5><div><ul><li><div>Th, 17.10.2024 10:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4359">Registration</a></div></li></ul></div><p>This webinar emphasizes the significance of capillary HPLC coupled with mass spectrometry.</p><h5><strong>14.&nbsp;Waters Corporation: Empower Tips: Ask Me Anything About Processing Data - Part 2</strong></h5><div><div><ul><li><div>Th, 17.10.2024 16:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/4360">Registration</a></div></li></ul></div><p>This is the second in our three-part Ask Me Anything webinar series on one of the most frequently asked about topics - processing data.</p></div><h5><strong>15.&nbsp;Waters Corporation: Nitrosamines</strong></h5><div><div><ul><li><div>Th, 17.10.2024 19:00 CEST</div></li><li><div><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/3445">Registration</a></div></li></ul></div><p>This webinar will address analytical challenges in the detection and control of nitrosamines and Nitrosamine Drug-Substance Related Impurities (NDSRIs).</p></div>

Webinars LabRulezLCMS Week 42/2024

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Webinars

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Science and research

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The Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences (IOCB Prague) is a leading scientific institution in the Czech Republic, recognized internationally. Its primary mission is basic research in the fields of chemical biology and medicinal chemistry, organic and material oriented chemistry, chemistry of natural compounds, biochemistry and molecular biology, physical chemistry, theoretical chemistry, and analytical chemistry. 

Ústav organické chemie a biochemie AV ČR

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LabRulez

Quantitative analysis of proteins (Bioanalytic) 

In our laboratory, proteomic analyses, including quantitative analysis of proteins, are performed by HPLC-MS or HPLC-MS/MS methods on a ZenoTOF 7600 high-resolution mass spectrometer (SCIEX).

<p><strong>Quantitative analysis of proteins is important in many industries and applications. Proteins include a diverse group of compounds that participate in virtually every biological process. Proteins are building blocks, hormones, transport molecules, cell receptors, cell growth and differentiation factors. Because of this, the analysis of protein interactions is crucial in drug design and production.</strong></p><p>Proteins are macromolecules - the basis of their structure are polypeptide chains containing from 100 to more than 10000 amino acids (10 - 100 kDa). Proteins are found in simple and complex forms. Complex proteins contain a non-protein component (sugar, lipids, metal ions or dyes) in their structure.</p><p>In our laboratory, proteomic analyses, including quantitative analysis of proteins, are performed by HPLC-MS or HPLC-MS/MS methods on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=ZenoTOF+7600"><strong>ZenoTOF 7600</strong></a> high-resolution mass spectrometer (SCIEX). We process the obtained results using Sciex OS, Spectronaut, Skyline software.</p><h4><strong>MS vs MS/MS</strong></h4><p>Quantitative analysis of intact protein (whole protein in intact form) leads to the concentration (determined from a prepared curve from a standard (with or without matrix) and confirmation of the identification by superimposing (on a mirrored basis) the MS spectrum of the protein determined in the sample to the MS spectrum of the standard. For your needs, we can generate a list of protein-structured compounds (impurities) that were identified in the sample and determine the ratio of their signal area to the signal area of the analyzed protein.</p><p>The result of proteomic quantitative peptide analysis allows precise and much more sensitive determination of protein concentration. The analysis is based on MS/MS data obtained for peptides obtained by digestion with proteolytic enzymes. The reported result consists of the concentration of the protein under study and confirmation of its presence based on the determined amino acid sequence. For such analyses, it is recommended to add standards internal standards.</p><blockquote><p><span style="font-size:22px;"><strong>Our offer:</strong></span></p><ul><li>For proteins and peptides with low masses - intact analysis with MS detection;</li><li>Proteins with masses &gt;20kDa require optimization of the digestion method and MS/MS detection. This results in specific signals from the analyzed protein in the mixture/matrix;</li><li>From complex matrices (including serum, cell culture, fungal suspension), sample analysis involves several steps:&nbsp;<ul><li>Sample purification;</li><li>Digestion to obtain unique peptides to distinguish the analyzed proteins from other proteins in the sample;</li><li>Sample preparation requires appropriate method optimization and validation (let's set validation criteria individually);</li></ul></li><li>Reported quantitative result includes confirmation of the presence of the protein in native form or a peptide derived from it (usually confirmation includes at least two peptide sequences);</li><li>Statistical analysis using tools: MarkerView (PCA), GraphPad Prism, Trends, Sciex OS.</li></ul><p><i>To optimize and validate the method, it is necessary to provide a standard, in the case of proteins with high masses - it is recommended to add an internal standard.</i></p><p>See also:</p><ul><li><a target="_blank" rel="noopener noreferrer" href="https://labexperts.com.pl/service/identyfikacja-bialek/"><strong>Protein identification</strong></a><strong>,</strong></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1296"><strong>Analysis and characterization of monoclonal antibodies</strong></a><strong>.</strong></li></ul></blockquote><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Quantitative_analysis_of_proteins_1_de494646f2.jpg" alt="Bioanalytic: Quantitative analysis of proteins." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Quantitative_analysis_of_proteins_1_de494646f2.jpg 200w," sizes="100vw" width="777" height="606"></p>

Identification of the substance – what is in your sample? (Bioanalytic)

We are able to identify metabolites and products of chemical reactions, contaminants of biological and chemical substances and preparations, and perform profiling of a series of samples to study the kinetics of transformations.

<h4><strong>Substance Identification - Looking for answers to what's in your sample? - we can help you.</strong></h4><p>Substance identification based on LC-MS/MS techniques can involve single compounds or mixtures of substances. In our analyses, we rely primarily on the knowledge and skills we have acquired after many years of work. For analysis, we use full laboratory facilities for sample preparation and high-resolution mass spectrometry (<a target="_blank" rel="noopener noreferrer" href="https://labexperts.com.pl/o-nas/">equipment</a>).</p><p>We use different work algorithms depending on the material we are dealing with and the analytical task to be solved. We use DDA (IDA) or DIA (SWATH) methods or targeted approaches (e.g. MRMhr) if we know what to expect. Thus, we are able to identify metabolites and products of chemical reactions, contaminants of biological and chemical substances and preparations, and perform profiling of a series of samples to study the kinetics of transformations.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Identification_of_the_substance_what_is_in_your_sample_2_0f425a648b.jpg" alt="Bioanalytic: Identification of the substance – what is in your sample" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Identification_of_the_substance_what_is_in_your_sample_2_0f425a648b.jpg 245w," sizes="100vw" width="1881" height="1156"></p><h4><strong>COST OF SERVICE</strong></h4><p>We determine the cost of the service individually. The final cost is influenced by many factors related to the nature of the sample itself and the client's expectations. Thus, the price depends, among other things, on the complexity of the sample preparation procedure, the scope and number of analyses, etc.</p><p><strong>The service includes (depending on individual arrangements):</strong></p><h5><strong>Development of sample preparation procedure</strong></h5><ul><li>Depending on the matrix and test substances.</li></ul><h5><strong>Development of analytical methods</strong></h5><ul><li>Application of high-resolution LC-MS/MS analyses using DDA (IDA) or DIA (SWATH) experiments and software to support the development of targeted screening methods.</li></ul><h5><strong>Analysis of results includes:</strong></h5><ul><li>finding and identifying the substances of interest, interpreting MS/MS spectra and proposing a structural formula.</li><li>Comparative analysis of sample series, e.g. PCA,</li><li>profiling of analytes in sample series</li></ul><h5><strong>Reporting of results</strong></h5><ul><li>Compile results into easy-to-read reports.</li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Identification_of_the_substance_what_is_in_your_sample_3_c1b8adc1d0.jpg" alt="Bioanalytic: Identification of the substance – what is in your sample" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Identification_of_the_substance_what_is_in_your_sample_3_c1b8adc1d0.jpg 245w," sizes="100vw" width="1919" height="1208"></p>

Characteristics of monoclonal antibodies (Bioanalytic)

At LabExperts, proteomic analyses are performed on a ZenoTOF 7600 high-resolution LC-MS/MS system (SCIEX). For MS/MS analyses, we use Collision induced dissociation (CID) and Electron-Activated Dissociation (EAD) fragmentation.

<p><strong>Antibodies are molecules made up of 4 polypeptide chains: two heavy (H - high-chain) linked to each other by a specific type of chemical bond - a disulfide bridge, and two light (L - light-chain) linked by an identical bond to the heavy chains. Monoclonal antibodies (mABs) are used in various forms - as free molecules, in the form of a drug-antibody combination, and in the form of an immunoconjugate, a drug-antibody-biopolymer combination.</strong></p><p>Their main use is in highly specific targeted therapy for cancer, cardiovascular, autoimmune and nervous system diseases. They are also used in the treatment of osteoporosis, Alzheimer's disease or transplantation. The range of applications of monoclonal antibodies is systematically expanded. The introduction of a new antibody to the market requires complex product characterization, which involves numerous proteomic analyses, which are performed by LC-MS/MS techniques, among others.</p><h4><strong>LC-MS/MS</strong></h4><p>The characterization of monoclonal antibodies involves many aspects, including, but not limited to, a suite of analyses related to confirmation of the structure, chain sequence and degree of modification of the antibody. Post-translational modifications (PTMs) are chemical changes that occur after the translation process. These changes generally result in an increase in the immunogenicity of the drug. For this reason, it is essential to monitor modifications as critical quality attributes (CQAs) during both the production and storage of the drug. To ensure monitoring of CQA points, the primary structure of the protein is characterized based on peptide mapping (peptide mapping) analysis. The protein under study is subjected to proteolytic digestion using enzymes such as trypsin, chymotrypsin, Lys-C, Glu-C. As a result of various proteolytic enzymes, a wide range of peptides is obtained. Ultimately, coverage of the entire sequence of the protein under study is obtained.</p><figure class="image"><img style="aspect-ratio:1536/947;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_3_d94fa1623b.jpg" alt="Bioanalytic: Characteristics of monoclonal antibodies." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_3_d94fa1623b.jpg 245w," sizes="100vw" width="1536" height="947"></figure><p>At LabExperts, proteomic analyses are performed on a <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=ZenoTOF+7600"><strong>ZenoTOF 7600</strong></a> high-resolution LC-MS/MS system (SCIEX). For MS/MS analyses, we use Collision induced dissociation (CID) and Electron-Activated Dissociation (EAD) fragmentation. This provides us with data for unambiguous identification of protein-building amino acids. We process the results using Sciex OS, Skyline and Spectronaut software.</p><blockquote><p><span style="font-size:22px;"><strong>We perform:</strong></span></p><ul><li>Whole protein analyses - Intact - to determine the exact mass and confirm the theoretical mass;</li><li>N-glycosylation profile determination (based on MS spectrum reconstruction):<ul><li>Glycosylated form;</li><li>Deglycosylated;</li><li>Glycosylated with reduced disulfide bridges;</li><li>Deglycosylated with reduced disulfide bridges.</li></ul></li><li>Peptide maps:<ul><li>Protein digestion with various proteases so that sequence coverage is close to 100%<ul><li>Analysis of PTMs:</li><li>Location of modifications</li></ul></li><li>[%] of modifications compared to the unmodified sequence</li></ul></li><li>TiPOS testing - (technical proof of similarity) - technical confirmation of the similarity of the protein synthesized by the customer (from several LOTs) with the target protein (provided as a template).</li></ul><p>In order to maintain the highest possible analytical standards, we perform monoclonal antibody analyses under the regime of an internally developed and implemented laboratory quality system based on GLP principles.</p><p>See also our other proteomics services: <a target="_blank" rel="noopener noreferrer" href="https://labexperts.com.pl/service/identyfikacja-bialek/"><strong>Protein identification</strong></a>, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1294"><strong>Protein quantitative analysis</strong></a></p></blockquote><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_2_cb93143fe6.jpg" alt="Bioanalytic: Bioanalytic Characteristics of monoclonal antibodies." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_2_cb93143fe6.jpg 245w," sizes="100vw" width="1536" height="908"></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_1_0e8ab7cdfa.jpg" alt="Bioanalytic: Bioanalytic Characteristics of monoclonal antibodies." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Characteristics_of_monoclonal_antibodies_1_0e8ab7cdfa.jpg 245w," sizes="100vw" width="1536" height="858"></p>

Analysis of Anabolic steroids (Bioanalytic)

We offer a highly specific and sensitive qualitative analysis of a wide profile of anabolics by LC-HR-MS/MS technique (detection limits range from 0.01 to 5 ng/ml, depending on the substance).

<h4><strong>Anabolic substances in urine</strong></h4><p>The effect of anabolic steroids (<a target="_blank" rel="noopener noreferrer" href="https://en.wikipedia.org/wiki/Anabolic_steroid"><strong>more information</strong></a>) is to stimulate the growth of muscle mass and sometimes also bone mass by stimulating protein synthesis or altering calcium metabolism. This effect is often used in sports doping. However, anabolic steroids also have many side effects, as they strongly affect the body's overall hormonal balance.</p><p>Anabolic steroids used by purely recreational athletes are still not banned in most countries, including Poland. Nevertheless, their amateur use during adolescence often results in the inability to continue a professional sports career into adulthood or leads to hormonal imbalances in the body.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_1_46fa2f41ca.jpg" alt="Bioanalytic: Analysis of Anabolic steroids." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_1_46fa2f41ca.jpg 245w," sizes="100vw" width="1200" height="740"></p><p>If you want to know whether and which anabolic substances are present in the samples of athletes, or you want to exclude doping in this area - use the offer carried out by us together with the company <a target="_blank" rel="noopener noreferrer" href="http://www.lab4tox.pl/"><strong>Lab4Tox</strong></a>. We offer a highly specific and sensitive qualitative analysis of a wide profile of anabolics by LC-HR-MS/MS technique (detection limits range from 0.01 to 5 ng/ml, depending on the substance). The use of high-resolution mass spectrometry (HR-MS and HR-MS/MS) in the analysis of anabolic steroids makes it possible to unequivocally confirm or exclude the presence of a given compound. The list of compounds at present includes:</p><ul><li><span class="ts-list-li-content">Boldenone undecylenate</span></li><li><span class="ts-list-li-content">Turinabol</span></li><li><span class="ts-list-li-content">Drostanolone enanthate</span></li><li><span class="ts-list-li-content">Drostanolone propionate</span></li><li><span class="ts-list-li-content">Methandienone</span></li><li><span class="ts-list-li-content">Methenolone acetate</span></li><li><span class="ts-list-li-content">Methenolone enanthate</span></li><li><span class="ts-list-li-content">Nandrolone phenylpropionate</span></li><li><span class="ts-list-li-content">Oxandrolone</span></li><li><span class="ts-list-li-content">Oxymetholone</span></li><li><span class="ts-list-li-content">Stanozolol</span></li><li><span class="ts-list-li-content">5-alpha-DHT</span></li><li><span class="ts-list-li-content">6-beta-hydroxymethandienone</span></li><li><span class="ts-list-li-content">19-norandrosterone</span></li><li><span class="ts-list-li-content">3'-hydroxystanazolol</span></li><li><span class="ts-list-li-content">4-beta-hydrozystanazolol</span></li><li><span class="ts-list-li-content">17-alpha-trenbolone</span></li><li><span class="ts-list-li-content">Testosterone caproate</span></li><li><span class="ts-list-li-content">Testosterone cypionate</span></li><li><span class="ts-list-li-content">Testosterone decanoate</span></li><li><span class="ts-list-li-content">Testosterone enanthate</span></li><li><span class="ts-list-li-content">Testosterone isocaproate</span></li><li><span class="ts-list-li-content">Testosterone phenylpropionate</span></li><li><span class="ts-list-li-content">Testosterone propionate</span></li><li><span class="ts-list-li-content">Testosterone undecanoate</span></li><li><span class="ts-list-li-content">Trenbolone acetate</span></li><li><span class="ts-list-li-content">Testosterone</span></li><li><span class="ts-list-li-content">Epitestosterone</span></li></ul><p><span class="ts-list-li-content">The list of compounds is not complete and will be systematically expanded according to customer needs.</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_2_78729119b0.jpg" alt="Bioanalytic: Analysis of Anabolic steroids is carried out by us together with the company Lab4Tox." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_2_78729119b0.jpg 245w," sizes="100vw" width="625" height="100"></p><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=54&amp;distributorItems=315"><span class="ts-list-li-content"><strong>Other analytical methods we offer</strong></span></a><span class="ts-list-li-content">.</span></p>

Analysis of Synthetic Peptide Hormones (Bioanalytic)

We offer highly specific and sensitive quantitative analysis of peptides by LC-MS/MS technique (LOQ < 1 ng/ml).

<h4><strong>Synthetic peptide hormones - a study of the purity of formulations and composition of dietary supplements.</strong></h4><p>In sports doping, small synthetic peptide hormones represent an increasingly attractive class of supplements for improving physical performance in amateur and professional athletes. Their use, depending on the type of peptide, is aimed primarily at building muscle tissue while intensifying fat reduction processes combined with the induction of the body's regenerative processes.</p><p>Do you want to be sure that the peptide you are using is in the product in the amount declared by the manufacturer? Or do you have doubts that the supplement you are using is contaminated with these compounds? If you are troubled by these or similar questions - feel free to contact us. We perform tests in cooperation with <a target="_blank" rel="noopener noreferrer" href="http://www.lab4tox.pl"><strong>Lab4Tox</strong></a>. We offer highly specific and sensitive quantitative analysis of peptides by LC-MS/MS technique (LOQ &lt; 1 ng/ml). At the moment, we analyze the following peptides:</p><ul><li><span class="ts-list-li-content">GHRP-2</span></li><li><span class="ts-list-li-content">GHRP-6</span></li><li><span class="ts-list-li-content">BPC-157</span></li><li><span class="ts-list-li-content">Melanotan-2</span></li></ul><p><span class="ts-list-li-content">The list of compounds is not complete and will be systematically expanded according to customer needs.</span></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_2_78729119b0.jpg" alt="Bioanalytic: Analysis of Synthetic Peptide Hormones. We perform tests in cooperation withLab4Tox." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Analysis_of_Anabolic_steroids_Bioanalytic_2_78729119b0.jpg 245w," sizes="100vw" width="625" height="100"></p><p><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=54&amp;distributorItems=315"><span class="ts-list-li-content"><strong>Other analytical methods we offer</strong></span></a><span class="ts-list-li-content">.</span></p>

Determination of nitrosamine content (Bioanalytic)

Our offer includes comprehensive development and validation of methods for determination of nitrosamines in pharmaceutical products. We perform the work according to the criteria and needs of the client based on the EMA note.

<p><strong>Determination of nitrosamines is one of the pharmaceutical industry's top priorities. After the presence of N-nitrosodimethylamine (NDMA) was first reported in an active pharmaceutical ingredient (API), the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) mandated the identification and control of all APIs and finished products in which there is a risk of nitrosamines.</strong></p><p>The content of nitrosamines in drug products can be controlled primarily by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis. More than that, a reliable analysis result in our offer consists of a well-chosen sample preparation procedure, a selective and sensitive analytical method, and the experience of the analyst, who selects the best method parameters at the optimization stage.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_2_ce4f440837.jpg" alt="Bioanalytic: Determination of nitrosamine content." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_2_ce4f440837.jpg 234w," sizes="100vw" width="1536" height="1024"></p><p><strong>We have gained experience working with leading pharmaceutical companies.</strong> As part of our ongoing work, we have developed analytical methods for small-molecule nitrosamines of 70-150 g/mol. These compounds are mainly formed during manufacturing processes and depend on the presence and type of excipients (e.g. NDBA - N-nitroso-dibutylamine). We have also developed methods for determining molecules with higher nitrosamine drug substance related impurities (NDSRIs). In our work, we focus on achieving the highest possible sensitivity, selectivity and precision. To this end, we individually select the parameters of the methods depending on the composition of the drug and the expected nitrosamines. Due to the above criteria and the required detection limits, we perform nitrosamine impurity determinations using LC-MS/MS kits based on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=QTRAP+5500%2B"><strong>QTRAP 5500+</strong></a> and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=QTRAP+6500%2B"><strong>QTRAP 6500+</strong></a> mass spectrometers (<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=55"><strong>SCIEX</strong></a>).</p><blockquote><p><span style="font-size:22px;"><strong>Our offer</strong></span></p><p><strong>Our offer includes comprehensive development and validation of methods for determination of nitrosamines in pharmaceutical products.</strong> <strong>We perform the work according to the criteria and needs of the client based on the EMA note.</strong></p><p>At the stage of sample preparation, we focus on:</p><ul><li>obtaining the <strong>highest possible recovery</strong> of nitrosamines (matrix purification);</li><li><strong>limiting additional synthesis</strong> of impurities (selection of appropriate equipment and reagents);</li><li><strong>ensuring the stability</strong> of the sample prepared for LC-MS/MS analysis.</li></ul><p>During the optimization of the LC-MS/MS method, we solve such problems as:</p><ul><li>selection of an <strong>appropriate internal standard</strong>;</li><li><strong>separation of the impurity from the API</strong>;</li><li>obtaining the expected <strong>sensitivity of the analytical method</strong>.</li></ul><p><strong>Quantitative analysis of nitrosamines requires sensitivity, precision and accuracy, which we provide to our customers.</strong></p></blockquote><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_1_d629e00e62.jpg" alt="Bioanalytic: Determination of nitrosamine content." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Determination_of_nitrosamine_content_Bioanalytic_1_d629e00e62.jpg 103w," sizes="100vw" width="1130" height="1710"></p>

Identification of the proteins (Bioanalytic)

We perform proteomic analyses on a state-of-the-art high-resolution ZenoTOF 7600 (SCIEX), in which the Zeno ion trap and EAD (Electron-Activated Dissociation) technology together make a powerful combination.

<p><strong>Protein identification is based on experimental measurement of peptide molecular weights and matching the amino acid sequence of the protein under study with protein sequences deposited in libraries (databases). Two approaches are most commonly used in proteomic studies: top-down and bottom-up. The top-down method involves analyzing full protein sequences. Bottom-up analyses, on the other hand, use digestion of proteins with proteolytic enzymes. The peptides obtained by digestion are analyzed using a mass spectrometer. Each analysis leads to a comparison of experimental data with data obtained by theoretical digestion of proteins described in the database.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Identification_of_the_proteins_Bioanalytic_2_6231b73f0d.jpg" alt="Bioanalytic: Identification of the proteins." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Identification_of_the_proteins_Bioanalytic_2_6231b73f0d.jpg 245w," sizes="100vw" width="1536" height="766"></p><h4><strong>LC-MS/MS</strong></h4><p>Tandem mass spectrometers are commonly used to determine protein sequences. They allow ion selection of pseudomolecular peptides and then fragmentation and measurement of the masses of the resulting fragments. Fragmentation using <i>collision induced dissociation</i>(CID ) is an invaluable tool in targeted and untargeted MS/MS analysis of small molecules. It is important to note that in proteomic analysis, data obtained using the aforementioned fragmentation method may leave gaps. For example, spectra obtained by CID fragmentation do not allow unambiguous differentiation of some amino acids: leucine from isoleucine (identical molar masses and CID fragmentation) and lysine from glutamine (very similar masses). For this reason, we perform proteomic analyses on a state-of-the-art high-resolution <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?manufacturerItems=52&amp;search=+ZenoTOF+7600"><strong>ZenoTOF 7600</strong></a> (SCIEX), in which the Zeno ion trap and EAD (<i>Electron-Activated Dissociation</i>) technology together make a powerful combination. It enables the creation of unique fragments to differentiate problematic amino acids and increase the sensitivity and specificity of the analysis.</p><blockquote><p><span style="font-size:22px;"><strong>We perform analyses of:</strong></span></p><ul><li>samples from solution;</li><li>samples from a gel;</li><li>samples from a complex matrix (by prior arrangement);</li><li>samples from lyophilizer;</li><li>samples previously prepared by the client.</li></ul><p>Qualitative analysis of proteins includes:</p><ul><li>To confirm the presence of a protein, the sample is subjected to digestion and the result obtained from experimental data is compared with theoretical data;</li><li>Comparison of experimental data on the masses of peptides included in the analyzed protein with the protein sequences described in the database;</li><li>Analysis and identification of the protein mixture;</li><li>Identification of post translational modifications (PTMs) (we use the tools: Skyline, Spectronaut).</li><li>Reporting of the result including, but not limited to, confirmation of the coverage of the protein sequence obtained in the test sample with the theoretical protein and the list of modified peptides, location and % of modifications.</li></ul><p><span style="font-size:22px;"><strong>Comparative analyses include:</strong></span></p><ul><li>Determining the composition of mixtures;</li><li>Comparison and profiling of sample series - protein identification and trend analysis.</li></ul><p><span style="font-size:22px;"><strong>Our offer also includes</strong>:</span></p><ul><li>Support in the planning and execution of studies (the scope is determined individually);</li><li>Assistance in the development of extraction methods from complex matrices - biological fluids, plant tissue, animal tissues (the scope is determined individually);</li><li>Statistical analysis using tools: MarkerView (PCA), GraphPad Prism, Trends, Sciex OS;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1294"><strong>Quantitative analysis of proteins</strong></a>;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1296"><strong>Analysis and characterization of monoclonal antibodies</strong></a>.</li></ul></blockquote><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Identification_of_the_proteins_Bioanalytic_1_97cf8c94b3.jpg" alt="Bioanalytic: Identification of the proteins." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Identification_of_the_proteins_Bioanalytic_1_97cf8c94b3.jpg 117w," sizes="100vw" width="1500" height="2000"></p>

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