Quantifying steroids in biological matrices has always presented analytical challenges. Complex structures, isobaric interferences, and matrix effects often limit accuracy and throughput — demanding a new approach beyond incremental improvements.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Advancing_Steroid_Quantification_Unlocking_Sensitivity_and_Selectivity_with_the_Thermo_Scientific_Stellar_Mass_Spectrometer_6e03ce7f59.jpg" alt="Thermo Fisher Scientific: Advancing Steroid Quantification: Unlocking Sensitivity and Selectivity with the Thermo Scientific Stellar Mass Spectrometer." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Advancing_Steroid_Quantification_Unlocking_Sensitivity_and_Selectivity_with_the_Thermo_Scientific_Stellar_Mass_Spectrometer_6e03ce7f59.jpg 245w," sizes="100vw" width="1024" height="512">The<a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1149"> Thermo Scientific Stellar MS</a> redefines how analysts approach steroid quantification, enabling rapid method optimization through advanced scanning flexibility, complementary fragmentation mechanisms, and enhanced selectivity.Rather than focusing on a single metric, this application note demonstrates how researchers can quickly develop more effective methods by leveraging the full capability of the Stellar MS to characterize compounds comprehensively in a single acquisition.<h3>A new paradigm for analytical quantitation</h3>Traditional LC-MS approaches often rely on incremental improvements in sensitivity or speed. Stellar MS instead provides a new analytical framework, designed to optimize every parameter of quantitative method creation.Key attributes include:<ol><li>PRM vs. SRM/MRM — improving instrument efficiency Parallel Reaction Monitoring (PRM) on Stellar MS provides comprehensive ion information for each target steroid, improving quantitation accuracy and efficiency over traditional MRM methods.</li><li>Complementary dissociation mechanisms — HCD and CID Stellar’s unique ability to combine beam-type (HCD) and resonant (CID) dissociation ensures full coverage of all 17 targeted steroids. This dual-mode approach effectively overcomes neutral water loss product ions — one of the key challenges in steroid analysis.</li><li>Next-level selectivity — MS² to MS³ transition The platform’s ability to perform MS³ experiments enhances selectivity and confidence, particularly in complex biological matrices, where interferences are common.</li><li>Product ion accumulation vs. dwell time — improved ion ratios and sensitivity By accumulating product ions rather than relying on fixed dwell times, Stellar MS achieves superior ion ratio precision and reproducibility — even at low spiking levels — without sacrificing throughput.</li></ol>&nbsp;<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Thermo_Fisher_Scientific_Advancing_Steroid_Quantification_Unlocking_Sensitivity_and_Selectivity_with_the_Thermo_Scientific_Stellar_Mass_Spectrometer_2_21e509451a.jpg" alt="Thermo Fisher Scientific: Advancing Steroid Quantification: Unlocking Sensitivity and Selectivity with the Thermo Scientific Stellar Mass Spectrometer: Figure 1. Representative EIC of 17 steroids in human serum using the Thermo Scientific Stellar mass spectrometer." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Thermo_Fisher_Scientific_Advancing_Steroid_Quantification_Unlocking_Sensitivity_and_Selectivity_with_the_Thermo_Scientific_Stellar_Mass_Spectrometer_2_21e509451a.jpg 245w," sizes="100vw" width="800" height="412"><h3>From characterization to confident method creation</h3>The power of Stellar MS lies in its comprehensive acquisition strategy. In this application note, a single injection per spiking level generated 125 distinct scan events, combining:<ul><li>HCD and CID MS² experiments</li><li>Targeted MS³ scans</li><li>Positive/negative polarity switching</li></ul>This workflow enables analysts to rapidly evaluate:<ol><li>Which dissociation mechanism produces the most informative spectra</li><li>Whether MS² or MS³ is best suited for each compound</li><li>The optimal product ions for confident post-acquisition quantitation</li></ol>By providing this complete dataset in a single run, Stellar MS dramatically accelerates method optimization and validation, supporting more efficient and reliable quantitation strategies.<h3>Driving method development efficiency</h3>The real strength of the Stellar MS isn’t just its speed or sensitivity — it’s the ability to experiment, compare, and decide in one workflow. Through simultaneous HCD/CID fragmentation, MS² and MS³ switching, and polarity control, users can determine the optimal analytical conditions across complex steroid panels quickly and confidently.This approach helps labs:<ul><li>Reduce development time</li><li>Maximize data quality</li><li>Improve long-term robustness and reproducibility</li></ul><h3>Conclusion</h3>The Stellar MS platform empowers analytical scientists to move beyond incremental performance metrics toward methodological innovation.By integrating comprehensive characterization and advanced scanning control, it enables faster, more confident decisions in quantitative steroid analysis — setting a new benchmark for flexibility and precision in LC-MS.Ask the Experts: On-Demand WebinarLearn more about how Stellar MS enhances steroid profiling and quantitative workflows. Watch the on-demand webinar where Thermo Fisher scientists discuss:<ul><li>Advanced method development strategies</li><li>Insights from the steroid panel dataset</li><li>Practical examples of leveraging Stellar’s flexibility for confident quantitation</li></ul><blockquote>Watch now: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/webinar/5117">Unlocking the Power of Quantitative Analysis with Stellar MS</a>Read the full application note: <a target="_blank" rel="noopener noreferrer" href="https://lcms.cz/paper/32415">Selective and sensitive measurement of 17 steroids in human serum using a Stellar mass spectrometer</a></blockquote><ul><li>Learn more about <a target="_blank" rel="noopener noreferrer" href="https://www.thermofisher.com/content/lifetech/global/en/home/industrial/mass-spectrometry/liquid-chromatography-mass-spectrometry-lc-ms/lc-ms-systems/orbitrap-lc-ms/stellar-mass-spectrometer/stellar-mass-spectrometer-applications.html">Stellar Mass Spectrometer applications</a></li><li>Visit us on&nbsp;<a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/showcase/72003132/admin/dashboard/">LinkedIn:</a>&nbsp;#MassSpectrometry #ClinicalResearch #LCMS #StellarMS</li></ul><blockquote><h5>Dane Walker</h5>Dane Walker is Regional Marketing Manager at Thermo Fisher Scientific, focused on LC-MS systems across EMEA. With a PhD in Surface Science and a background in scientific instrumentation, he blends technical expertise with strategic marketing to drive data-led campaigns. Passionate about communicating science and enabling innovation, Dane enjoys powerboating, cycling, stargazing, and talking endlessly about mass spectrometry.</blockquote>

Advancing Steroid Quantification: Unlocking Sensitivity and Selectivity with the Thermo Scientific Stellar Mass Spectrometer

<h4>The problem&nbsp;</h4>We live in a “global village” but we are all different and we use many different languages to communicate.&nbsp;Even when people speak the same language, the same words may be used with different meanings. See, for example, some different uses of the word “standard” in English – a normative document, a solution of known concentration or even a type of flag.&nbsp;Even in the same area of activity misunderstandings do occur which may result in a waste of time and money, or even worse consequences.&nbsp;When talking about laboratory tests, lack of a full understanding of key terms and inappropriate translations leaves room for different interpretations of stated requirements. For example, there have historically been a number of different but related uses of the term ‘detection limit’. This may lead to inadequate fulfilment of those requirements, thus becoming an obstacle to the production of analytical data that is fit for its intended use. A shared interpretation of concepts related to quality in measurement between laboratories and accreditation bodies is also crucial for a fair and harmonised approach to laboratory assessment across the world.&nbsp;<h4>A common language&nbsp;</h4>We need a common language; clear and unambiguous. We need a common vocabulary to provide consistent definitions of concepts with their associated terms. To this end, several organisations participate in a joint effort to develop and maintain an international vocabulary of metrology (VIM) [1], which aims to achieve a unique understanding of concepts related to measurements across all sectors. The VIM is a normative reference in the standards ISO/IEC 17025, ISO 15189 and ISO/IEC 17043, and is therefore a key document for all organisations seeking accreditation.&nbsp;<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Eurachem_You_talk_we_understand_The_way_out_of_the_tower_of_Babel_Standards_8d947dbd5f.jpg" alt="Eurachem: You talk, we understand – The way out of the tower of Babel_A common language" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Eurachem_You_talk_we_understand_The_way_out_of_the_tower_of_Babel_Standards_8d947dbd5f.jpg 245w," sizes="100vw" width="1126" height="499"><h4>Is this enough?&nbsp;</h4>For laboratory staff some problems still stand and need to be addressed:&nbsp;<ul><li>Many people are confused about both concepts and terms.&nbsp;</li><li>It may be difficult to understand the “formal” VIM definitions – they are short statements intended to apply to many different measurement sectors.</li><li>Translations into local languages may add to the confusion, e.g. if different terms are used in different sectors for the same VIM concept and the translator overlooks this issue.</li><li>Some substantial changes to terminology have been made in an attempt to accommodate chemical and biological measurements. Some “old” concepts still stand but have new names, e.g. “within laboratory reproducibility” vs. “intermediate precision”.&nbsp;</li></ul>To address these issues Eurachem has produced a guide [2] which provides an accessible reference containing simple and in context explanations of terms most relevant to analysts, with additional examples relating those concepts to chemical and biological measurements.&nbsp;<h4>Do you understand the meaning of these terms?&nbsp;</h4>Are you interested in a quantity or a quality? Depending on the answer, will you use a measurement procedure or an examination? Is method verification the same or less demanding than method validation? Do you need a calibration or just performance verification? Do you have a measurement standard in your laboratory? Is metrological traceability something you worry about? These are examples of the terms supporting the reliability of any measurement result that need to be clearly understood.&nbsp;<h5>Example – Accuracy, trueness, precision: are they synonyms?&nbsp;</h5><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Eurachem_You_talk_we_understand_The_way_out_of_the_tower_of_Babel_accuracy_231e1e005e.jpg" alt="Eurachem: You talk, we understand – The way out of the tower of Babel_Example – Accuracy, trueness, precision" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Eurachem_You_talk_we_understand_The_way_out_of_the_tower_of_Babel_accuracy_231e1e005e.jpg 146w," sizes="100vw" width="754" height="807"><ul><li>Closeness between measurement results (relating to the concept of precision)</li><li>Closeness of the average of the measurement results to a reference value (relating to the concept of trueness)&nbsp;</li><li>Closeness of an individual measurement result to the true value (relating to the concept of accuracy)&nbsp;</li></ul>The different situations in the figure show the effect of:&nbsp;<ol style="list-style-type:lower-latin;"><li>poor precision and trueness (imprecise and biased)&nbsp;</li><li>good precision, poor trueness (precise but biased)&nbsp;</li><li>poor precision, good trueness (imprecise but unbiased)&nbsp;</li><li>good precision and trueness (precise and unbiased)&nbsp;</li></ol>Improving both precision and trueness is expected to improve accuracy.&nbsp;<h4>Is all this relevant to you?&nbsp;</h4>Do you work in a laboratory, either as an analyst, quality manager or director? Are you involved in other activities related to analytical work, such as providing proficiency tests, producing reference materials or accrediting any such activity? Do you teach or study subjects related to measurements? Do you use measurement results or prescribe how to use them? If the answer is yes to any of these questions, the following reference documents are also for you.

You talk, we understand – The way out of the tower of Babel

Visit the LabRulez booth at <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/events/871">analytica 2026</a> in Munich, 24. - 27. March 2026! You can find us in Hall A1, Booth A1.506. At the booth, you will be able to explore live demonstrations of how the LabRulez portals work and see selected analytical instruments from our pre-owned marketplace section.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_at_Analytica_2026_Floor_plan_55e1057e03.jpg" alt="LabRulez at Analytica 2026: Hall A1 Floor Plan and Booth Location" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_at_Analytica_2026_Floor_plan_55e1057e03.jpg 245w," sizes="100vw" width="1516" height="881"><h3>Would you like to learn how the LabRulez portals work?</h3>➡︎ <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/">LabRulezLCMS</a> ➡︎ <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/">LabRulezGCMS</a> ➡︎ <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/">LabRulezICPMS</a><ul><li>A unique online solution for your marketing</li><li>A targeted audience from the analytical chemistry community</li><li>Modern technologies and a unique concept</li><li>Design optimized for most smart devices – PC, mobile, and tablet</li><li>Advanced statistical tools fully available to users</li></ul><h3>Services Offered</h3><h4>News + Company Updates</h4><ul><li>Publish news related to your instrumentation, applications, services, events, job openings, and more</li><li>News is shared both on our portals and across our social media channels</li></ul><h4>Library – All Documents in One Place</h4><ul><li>Applications, posters, presentations, technical articles, brochures, specifications</li><li>Fast and advanced search capabilities</li><li>Over 32,000 documents currently available in our database</li></ul><h4>Webinars</h4><ul><li>Upcoming and on-demand webinars from the field of analytical chemistry</li><li>More than 5,500 webinars covering chromatography, mass spectrometry, and spectroscopy</li></ul><h4>Careers</h4><ul><li>Targeted job listings in analytical chemistry</li><li>Clear filtering by specialization or location</li><li>Increased visibility for job postings</li></ul><h4>Companies</h4><ul><li>Company profile with key information and contacts</li><li>Integration of all your activities on our portal: News, Library documents, Webinars, Upcoming Events, Careers, and mentions by partners or customers</li><li>Connection to your Products listed on LabRulez portals</li></ul><h4>Products and Services</h4><ul><li>Comprehensive overview of instrumental techniques and services</li><li>Targeting the analytical chemistry community and customers in the growing online environment</li></ul><h4>Marketplace</h4><ul><li>Buying and selling pre-owned analytical equipment</li><li>We purchase analytical instrumentation of all brands and ages</li><li>Sell your equipment through LabRulez</li><li>Request and purchase complete systems, modules, or spare parts with warranty and service</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Lab_Rulez_at_analytica_2026_bazar_7edf4a0872.jpg" alt="LabRulez at analytica 2026" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Lab_Rulez_at_analytica_2026_bazar_7edf4a0872.jpg 88w," sizes="100vw" width="1600" height="2844"><h3>What Will Be on Display at Our Booth?</h3>Come and explore selected spare parts and modules from our marketplace section. These include both new, tested, and pre-owned items.<h4><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar">LabRulezLCMS Marketplace</a></h4>Spare parts for LC/MS:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=378">Vacuum turbo pumps for LC/MS</a></li></ul>Spare parts for HPLC:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=397">Degassers for HPLC/UHPLC</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=368&amp;search=valve">Automatic valves for HPLC</a></li><li>HPLC components for Agilent 1100, 1200, and 1260 Infinity II series</li></ul>Consumables:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=377">HPLC lamps</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/bazaar?category=48">HPLC columns</a></li></ul><h4><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar">LabRulezGCMS Marketplace</a></h4>Spare parts for GC/MS:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=367&amp;search=turbo">Turbo pumps for GC/MS</a></li></ul>Spare parts for GC:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=366&amp;search=EPC">EPC modules</a></li></ul>Sample injection<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar/432">Autosampler 7693A for GC</a></li></ul>Consumables:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/bazaar?category=7">GC columns </a>and selected accessories</li></ul>

LabRulez at analytica 2026

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Eurachem is a European network promoting international traceability of chemical measurements and good analytical practice. It provides expert guidance, supports method validation, uncertainty evaluation, proficiency testing, and facilitates discussion on technical and policy issues in analytical quality.

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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).

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.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.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">ZenoTOF 7600</a> high-resolution mass spectrometer (SCIEX). We process the obtained results using Sciex OS, Spectronaut, Skyline software.<h4>MS vs MS/MS</h4>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.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.<blockquote>Our offer:<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>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.See also:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://labexperts.com.pl/service/identyfikacja-bialek/">Protein identification</a>,</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1296">Analysis and characterization of monoclonal antibodies</a>.</li></ul></blockquote><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">

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>Substance Identification - Looking for answers to what's in your sample? - we can help you.</h4>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>).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.<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"><h4>COST OF SERVICE</h4>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.The service includes (depending on individual arrangements):<h5>Development of sample preparation procedure</h5><ul><li>Depending on the matrix and test substances.</li></ul><h5>Development of analytical methods</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>Analysis of results includes:</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>Reporting of results</h5><ul><li>Compile results into easy-to-read reports.</li></ul><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">

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.

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.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.<h4>LC-MS/MS</h4>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.<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>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">ZenoTOF 7600</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.<blockquote>We perform:<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>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.See also our other proteomics services: <a target="_blank" rel="noopener noreferrer" href="https://labexperts.com.pl/service/identyfikacja-bialek/">Protein identification</a>, <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1294">Protein quantitative analysis</a></blockquote><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"><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">

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>Anabolic substances in urine</h4>The effect of anabolic steroids (<a target="_blank" rel="noopener noreferrer" href="https://en.wikipedia.org/wiki/Anabolic_steroid">more information</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.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.<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">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/">Lab4Tox</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:<ul><li>Boldenone undecylenate</li><li>Turinabol</li><li>Drostanolone enanthate</li><li>Drostanolone propionate</li><li>Methandienone</li><li>Methenolone acetate</li><li>Methenolone enanthate</li><li>Nandrolone phenylpropionate</li><li>Oxandrolone</li><li>Oxymetholone</li><li>Stanozolol</li><li>5-alpha-DHT</li><li>6-beta-hydroxymethandienone</li><li>19-norandrosterone</li><li>3'-hydroxystanazolol</li><li>4-beta-hydrozystanazolol</li><li>17-alpha-trenbolone</li><li>Testosterone caproate</li><li>Testosterone cypionate</li><li>Testosterone decanoate</li><li>Testosterone enanthate</li><li>Testosterone isocaproate</li><li>Testosterone phenylpropionate</li><li>Testosterone propionate</li><li>Testosterone undecanoate</li><li>Trenbolone acetate</li><li>Testosterone</li><li>Epitestosterone</li></ul>The list of compounds is not complete and will be systematically expanded according to customer needs.<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"><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?category=54&amp;distributorItems=315">Other analytical methods we offer</a>.

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.

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.<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"><h4>LC-MS/MS</h4>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 collision induced dissociation(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">ZenoTOF 7600</a> (SCIEX), in which the Zeno ion trap and EAD (Electron-Activated Dissociation) 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.<blockquote>We perform analyses of:<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>Qualitative analysis of proteins includes:<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>Comparative analyses include:<ul><li>Determining the composition of mixtures;</li><li>Comparison and profiling of sample series - protein identification and trend analysis.</li></ul>Our offer also includes:<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">Quantitative analysis of proteins</a>;</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1296">Analysis and characterization of monoclonal antibodies</a>.</li></ul></blockquote><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">

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