Written by: Jenna Kerry, MSc Published: November 6, 2024 Contents Why is Antibody Humanization Important? What is Antibody Humanization? Antibody Humanization Techniques Current Challenges of Antibody Humanization The Future of Human Antibody Therapeutics Fully Human Antibody Discovery / Human Antibody Discovery From Human Serum Why is Antibody [...]
Written by: Genya Gorshtein, MSc Published: October 16, 2024 Contents Introduction Epitope Mapping Methods Addressing the Limitations of HDX-MS for Epitope Mapping HDX-MS as a Stand Alone Method for Epitope Mapping Epitope Mapping with HDX-MS at Rapid Novor Introduction Understanding antibody-antigen interactions is critical for [...]
In this webinar, you will learn: How mAb irreproducibility could unknowingly impact your research outcomes Proven strategies to mitigate reproducibility risks when using research reagents The role of antibody sequencing in ensuring reproducibility with mAbs Rapid, cost-effective sequence confirmation for routine applications The next frontier: polyclonal antibody sequencing and recombinant pAbs [...]
Written by: Jenna Kerry, MSc Published: September 17, 2024 Contents Introduction Agonistic Antibody Mechanism of Action Agonistic Antibody Development Challenges Optimizing Agonistic Antibodies Agonistic Antibody Development Introduction In the last decade, immunotherapy has rapidly transformed treatments for cancer, autoimmune disorders, and infectious diseases. While the [...]
Written by: Genya Gorshtein, MSc Published: August 26, 2024 Contents Introduction Advantages of Antibody Discovery In Diverse Species Unique Features of Antibodies from Diverse Hosts Antibody Discovery Platforms for Diverse Hosts Introduction The historical use of mice for monoclonal antibody generation has been driven [...]
Written by: Jenna Kerry, MSc Published: August 14, 2024 Contents What are Antagonistic Antibodies? How do Antagonistic Antibodies Work? Antagonistic Antibody Opportunities Antagonistic Antibody Obstacles Antibody Discovery Platform for Antagonistic Antibodies What are Antagonistic Antibodies? Antagonistic antibodies are a monoclonal antibody that bind to and [...]
Written by: Genya Gorshtein, MSc Published: August 8, 2024 Contents Introduction What are Protein Sequence Variants Impact of Sequence Variants on Monoclonal Antibodies Challenges in Sequence Variant Analysis and Detection Sequence Variant Analysis Service Introduction Sequence variants are unwanted impurities in antibody and biotherapeutic [...]
Written by: Jenna Kerry, MSc Published: August 1, 2024 Contents Introduction Disulfide Bond Structure and Function in Monoclonal Antibodies Disulfide Scrambling in Monoclonal Antibodies Impact of Disulfide Bonds on Monoclonal Antibodies Strategies to Prevent Disulfide Scrambling in Monoclonal Antibodies Disulfide Bond Analysis for Monoclonal Antibodies Introduction [...]
Written by: Vanessa Yoon Calvelo, PhD Published: June 6, 2023 Updated: July 7, 2024 Contents Introduction Early Discovery and Development of Therapeutic Antibodies Target Identification and Validation Antibody Discovery and Expression Lead Characterization and Selection Lead Engineering and Optimization Candidate Selection and Downstream Characterization Introduction [...]
Written by: Jenna Kerry, MSc Published: June 10, 2024 Contents Introduction Advantages of Recombinant Antibody Cocktails Recombinant Antibody Cocktail Formats Engineering Recombinant Antibody Cocktails Production of Recombinant Antibody Cocktails Recombinant Antibody Cocktail Applications Generate Antibody Cocktails with De Novo Antibody Sequencing Service Introduction Recombinant antibody [...]
Written by: Jenna Kerry, MSc Published: May 1, 2024 Contents What are Autoantibodies? How are Autoantibodies Produced? How do Autoantibodies Become Pathogenic? Triggers for Pathogenic Autoantibodies Autoantibodies in Diagnosis and Treatment Autoantigen Discovery and Characterization Gaining Insight into Autoantibodies What Are Autoantibodies? Autoantibodies are a [...]
Written by: Genya Gorshtein, MSc Published: April 19, 2024 Contents Introduction Bispecifics and Multispecific Antibody Formats Mechanism of Action of Bispecific Antibodies Bispecific and Multispecific Development with Rapid Novor Introduction The complex nature of diseases often limits the therapeutic efficacy of monovalent antibodies. To [...]
Written by: Genya Gorshtein, MSc Published: April 3, 2024 Contents Introduction Membrane Proteins are Difficult Antigens Strategies for Utilizing Membrane Proteins as Antigens in Antibody Discovery In Vivo Antibody Discovery Using Mass Spectrometry Introduction Membrane proteins orchestrate a wide range of cellular functions, such [...]
Written by: Jenna Kerry, MSc Published: March 5, 2024 Contents Introduction to Flow Cytometry Understanding Flow Cytometry Interpreting Flow Cytometry Data Antibody Selection for Flow Cytometry Antibody Validation for Flow Cytometry Beyond Binding Assays Introduction to Flow Cytometry Flow cytometry is used by a wide [...]
Written by: Jenna Kerry, MSc Published: February 2, 2024 Contents Introduction Recombinant Expression Systems Recombinant Expression of Polyclonal Antibodies Rapid Novor's Services Introduction Once antibody discovery, lead selection, and characterization is complete, researchers have a range of expression systems to choose from to produce the antibody [...]
Written by: Genya Gorshtein, MSc Published: January 8, 2024 Contents Introduction Glycoengineering of Therapeutic mAbs Glycoengineering Methods Informed Glycan Engineering through LC-MS Glycan Analysis Introduction Glycosylation plays a crucial role in antibody functions, influencing protein folding, trafficking, stability, and half-life. While the glycosylated residue [...]
Written by: Jenna Kerry, MSc Published: November 29, 2023 Contents Introduction Nanobody Structure Generation and Production Therapeutic and Diagnostic Applications Nanobody De Novo Sequencing Introduction Nanobodies, also known as single-domain antibodies or VHHs, are derived from the immune systems of camelids and are heavy-chain-only antibodies. Despite [...]
Written by: Jenna Kerry, MSc Published: October 25, 2024 Published: October 25, 2023 Contents Introduction What is Affinity Maturation? How Does Somatic Hypermutation Work? Applications of Affinity Maturation De Novo Proteomic Sequencing of Antibodies Introduction The immune system is a complex system responsible for [...]
Written by: Genya Gorshtein, MSc Published: October 18, 2023 Contents Introduction Types of Glycans How are Proteins Glycosylated? Why are Proteins Glycosylated? Protein Glycosylation in the Immune System Importance of Understanding Glycan Profiles of Proteins Discover Glycan Sites and Profiles with Rapid Novor Introduction [...]
Written by: Genya Gorshtein, MSc Published: September 12, 2023 Contents Introduction Mechanisms That Influence Protein Aggregation and Stability Sequence and Structural Characteristics That Impact Antibody Aggregation and Stability Engineered Antibody Formats Influence Protein Aggregation Strategies to Reduce Antibody Aggregation Propensity Improving Antibody Developability with Next Generation Protein Sequencing and Proteomics [...]
Written by: Genya Gorshtein, MSc Published: June 13, 2023 Contents Introduction How do Primary and Secondary Antibodies Work? How are Primary and Secondary Antibodies Generated? Selecting Primary and Secondary Antibodies for Immunoassays Developing Reliable Reagents for Immunoassays with Rapid Novor Introduction Immunoassays have a [...]
Written by: Genya Gorshtein, MSc Published: May 9, 2023 Contents Introduction Hybridoma Instability Leads to mAb Irreproducibility Batch-to-Batch Variation in Polyclonal Antibodies Antibody Sequencing and Recombinant Expression Ensures Reproducible Antibody Reagents Generating Reproducible Reagents with Rapid Novor Introduction Antibodies (Abs) are indispensable tools in [...]
Written by: Vanessa Yoon Calvelo, PhD Published: April 24, 2023 Contents Introduction Functions of Therapeutic Antibodies Functional Assays for Therapeutic Antibodies Functional Characterization in the Therapeutic Antibody Discovery Process Introduction Monoclonal antibodies (mAbs) and related biological products often present as ideal therapeutics largely due to: Their [...]
Written by: Genya Gorshtein, MSc Published: April 21, 2023 Contents Introduction Target Specification for First Generation Antibodies Patent Strategies for Second-Generation Antibodies Next Generation Protein Sequencing and Proteomics Strengthens IP Protections Introduction Intellectual property (IP) protection is a critical step during the commercialization of antibodies [...]
Written by: Vanessa Yoon Calvelo, PhD Published: April 14, 2023 Contents Introduction Immunochemical Properties of Therapeutic Antibodies Rapid Immunochemical Characterization of Antibodies Introduction Therapeutic antibodies, predominantly monoclonal antibodies (mAbs), are a rapidly expanding class of drugs with over 100 mAb-based biologics now approved for the treatment [...]
In This Webinar, You Will Learn: Why IP is important in the development of antibody-based assets Patent application process and requirements How to strengthen patent applications Applications of De Novo Protein Sequencing, SPR assays, and HDX-MS for antibody-related patent applications Abstract In the development of novel and innovative antibody-based assets, [...]
Written by: Genya Gorshtein, MSc Published: February 17, 2023 Contents The Drug Development Pathway Target Discovery and Validation Antibody Discovery Characterization and Lead Selection Engineering and Optimization Candidate Selection Therapeutic Antibody Discovery with Rapid Novor The Drug Development Pathway The four phases in the drug [...]
Written by: Genya Gorshtein, MSc Published: January 6, 2023 Contents Introduction What is Affinity? What is Avidity? Measuring Affinity and Avidity: ELISA or SPR? Measuring Affinity And Avidity For Biological Applications Affinity and Avidity Analysis via SPR with Rapid Novor Introduction Kinetics of antibody-antigen interactions [...]
Written by: Genya Gorshtein, MSc Published: November 1, 2022 Contents Introduction Approaches for Engineering Antibody Therapeutics Driving Antibody Engineering with Next Generation Protein Sequencing and Proteomics Introduction Antibody engineering encompasses various development, production strategies, and modification techniques to improve the biological properties of monoclonal antibodies (mAbs) [...]
Biological processes are driven by molecules that interact through specific molecular contacts, often to form a stable complex. These interactions are typically defined by the principles of thermodynamics as well as biomolecular structure and recognition. At the simplest level is the interaction between a target molecule with a specific binding site and a probing molecule that binds to that site, resulting in the bound complex.
Written by Genya Gorshtein, MSc Updated: January 27, 2023 (Published: August 31, 2022) Contents How is Antibody Diversity Generated? Which Steps Contribute to the Generation of Antibody Diversity? Antibody Loci and V(D)J Recombination Somatic Hypermutation Class Switch Recombination De Novo Proteomic Sequencing of Antibodies How is Antibody [...]
Written by: Genya Gorshtein, MSc Published: August 18, 2022 Contents Introduction How are Polyclonal Antibodies Produced? Applications of Polyclonal Antibodies Challenges of Polyclonal Antibodies De Novo Polyclonal Antibody Sequencing Introduction Polyclonal antibodies (pAbs) are a heterogeneous mix of antibodies derived from B cells in the [...]
Written by: Vanessa Yoon Calvelo, PhD Updated: January 19, 2023 (Published: August 11, 2022) Contents What is Surface Plasmon Resonance Spectroscopy? What is SPR Used For? How Does SPR Work? SPR Experimental Workflow SPR Sensorgram SPR Advantages SPR Applications SPR Antibody-Antigen Interaction Analysis at Rapid Novor What is [...]
Written by Genya Gorshtein, MSc August 10, 2022 Contents General Structure of Antibodies Fab and Fc Fragments Hinge Region CDR and FR Regions Antibody Structure Analysis Services General Structure of Antibodies Antibodies or immunoglobulins (Ig) maintain a common quaternary structure consisting of two identical heavy chains (HCs) and [...]
Written by: Vanessa Yoon Calvelo, PhD Published: August 3, 2022 Contents What are Biosimilar Drugs? Why are Biosimilars Being Developed? Biosimilars are not the Equivalent of Generics Biosimilar Development Biosimilar Monoclonal Antibodies De Novo Protein Sequencing Solutions in Biosimilar Development What are Biosimilar Drugs? Biosimilar drugs, [...]
Characterization of proteins and protein complexes is a major keystone of structural biology. As our understanding of cellular processes continues to evolve from simple pathways to complicated networks, our need for advanced analytical methods is quite apparent. Mass spectrometry (MS)-based structural approaches can be used to study protein conformational changes and dynamics, protein motion/flexibility, ligand-protein binding, and protein-protein interfaces.
Written by: Vanessa Yoon Calvelo, PhD Published: July 11, 2022 Contents What is Gene Therapy? What are Adeno-Associated Viruses? Engineering of AAVs for Gene Therapy Engineering AAVs for Improved Transduction Engineering AAVs for Improved Immunogenicity De Novo Protein Sequencing Applications in AAV Characterization and Development What is Gene [...]
Written by: Vanessa Yoon Calvelo, PhD Published: June 13, 2022 Contents What is CAR-T Cell Therapy? CAR Structure and Function CAR-T Cell Development Engineering Strategies for CAR-T Cells De Novo Protein Sequencing Applications in CAR-T Cell Development What is CAR-T Cell Therapy? The infusion of T cells [...]
Written by: Vanessa Yoon Calvelo, PhD Updated: January 19, 2023 (Published: June 2, 2022) Contents What are post-translational modifications (PTMs)? Impact of PTMs Types of PTMs PTMs increase microheterogeneity of antibodies PTM Analysis by LC-MS The Importance of Post-Translational Modifications (PTMs) Post-translational modifications (PTMs) are processing [...]
Written by: Yuning Wang, PhD Updated: January 26, 2023 (Published: June 3, 2022) Contents Introduction The Four Levels of Protein Structure How are Protein Structures Studied? Introduction Structural information provides a great deal of understanding of how a protein works, which can allow us to [...]
The most straightforward solution would be to determine sequences of the dominating antibody forms in a polyclonal mixture to enable recombinant antibody generation and ensure reproducibility. This was recently made possible by the development of polyclonal antibody sequencing technology, which will be reviewed in this article.
The origin of hydrogen-deuterium exchange (HDX) dates back to the 1950s, when protein scientist Linderstrøm-Lang created a method involving protein deuteration to distinguish amide hydrogens participating in secondary structures. Today, scientists frequently rely on HDX data to investigate protein structure, conformational dynamics, and protein-ligand interaction.
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
Written by: Yuning Wang, PhD Updated: January 18, 2023 (Published: January 21, 2022) Contents Discovery of Camelid Antibodies What are Camelid Antibodies? Structure of Camelid Antibodies and Nanobodies Advantages of Camelid Antibodies and Nanobodies Camelid Antibodies and Nanobodies for Therapeutic and Research Applications How are Camelid Antibodies [...]
Recombinant antibodies are artificially synthesized antibodies. Recombinant antibodies are generated from expression systems (e.g., E.coli, yeast, mammalian cell lines) via transfection with two separate plasmids encoding the amino acid sequences for the light and heavy chains, respectively. In order to recombinantly produce mAbs, the amino acid sequence of the light and heavy chains must be known. There are many ways to obtain the sequence of an antibody.
Since 2006, the One Health Initiative (OHI)’s goal has been to demonstrate the inextricable link between humans, animals, and the environment. Certainly, the current global pandemic is a great testament to the ties between climate change, humans, and animals that OHI has been working to highlight. The rise of other zoonotic diseases (e.g., Hendra, and Nipah viruses) not only directly affect humans through disease transmission but may also result in deep impacts to the food supply
Written by Yuning Wang, PhD and María Gerpe, PhD November 12, 2021 Contents IgBLAST Definition Why was IgBLAST Created? The Functions of IgBLAST How to Use IgBLAST Limitations of IgBLAST Additional Resources IgBLAST Definition Developed by the National Center for Biotechnology Information (NCBI), IgBlast is [...]
Peptide mapping is a widely used analytical technique to verify the primary structure (amino acid sequence) and characterize the chemical modifications of a protein. It analyzes peptides generated from the digestion of an isolated protein, or a protein mixture
Amino acids are small organic molecules that make up peptides and proteins. All living organisms share the same set of amino acids. Amino acids come together in different orders (sequences) to form proteins. As such, each type of protein has a different three-dimensional structure and biological activity.
Our team, along with four other industry panelists, discuss ways to safeguard their research through recombinant antibodies, cell culturing procedures, antibody protein sequences, and reference identifiers.
The transition from polyclonal antibody drugs to a more targeted monoclonal approach was made possible through a series of scientific and technological advancements; the most notable of which is the hybridoma technique developed by Köhler and Milstein, which allowed the generation of pure antibodies at scale.
Protein sequencing is a method that typically utilizes mass spectrometry (MS) to determine the amino acid code of a protein1. Prior to the development of mass spectrometry, Edman degradation, a method involving the stepwise degradation of peptides to derive the order of amino acids, was the mainstream approach. Nowadays, mass spectrometry is favored due to its ease of use and high throughput capabilities, though Edman degradation is still employed for specific applications in which the technique is well suited.
Monoclonal antibodies (mAbs) are homogenous antibodies that bind to a single epitope on an antigen. Kohler and Milstein generated the first mAbs when they developed hybridoma technology in the 1970s. Because of the specificity, homogeneity and unlimited availability, mAbs are valuable reagents used in a variety of important applications including treatment and diagnosis of diseases
Amino acids (aa)—the building blocks of proteins—are simple molecules characterized by a variable R group flanked either side by an amino group and a carboxyl group. With around 20 different commonly found amino acids, each one can bond with another to produce chains that can be classified as peptides (typically below 50 aa) and proteins (sequences above 50 aa)—molecules ubiquitous to every known organism.
Amino acid sequencing is commonly performed using Edman degradation or mass spectrometry (MS). While mass spectrometry is favoured for its high throughput capabilities and ease of use, both techniques possess their own features and limitations. This article summarizes some of the key pain points inherent in the two methodologies when determining the amino acid sequence.
Antibodies are used in a variety of ways in academia and industry, from tools to therapeutics. Because antibodies are produced using live processes, which are naturally error-prone, validation is required from time to time. Furthermore, to develop biological therapeutics, the protein sequence must be confirmed as part of the regulatory process.
DNA sequencing is the process of determining the precise order of four nucleotides bases—adenine (A), guanine (G), cytosine (C), and thymine (T)—that make up the DNA molecule. From Sanger sequencing to next-generation sequencing (NGS), DNA sequencing’s accessibility and ease of use make it one of the most widely used technologies in life sciences.
As proteins are assembled, they fold into different structural orders: from primary to quaternary. The exact sequence of the primary structure (the amino acid sequence) will dictate how a protein will fold and function. The importance of the primary structure has been noted in several studies, where changes in the original amino acid sequence have resulted in affinity problems, binding disruption, reduced half-life, and higher aggregation odds.
Proteins are composed of peptide chains, which in turn are made up of a string or linear sequence of amino acids. Every amino acid has a basic structure containing an amino (-NH2) group and a carboxylic (-COOH) group (Figure 1B). To form a peptide, amino acids link to each other via a peptide bond, which involves the reaction between the carboxylic group of one amino acid and the amine group of another amino acid.
Protein mass spectrometry refers to the use of mass spectrometry in studying and characterizing proteins, including their quantification, profiling, interaction mapping, and identification of their post-translational modifications. Protein mass spectrometry may also be referred to as mass spectrometry-based proteomics. Mass spectrometry-based proteomics comprises three approaches: top-down, middle-down, and bottom-up proteomics.
This webinar offers insight into how DNA and protein sequencing compare to each other.
Antibody sequences are critical for antibody engineering and protein characterization in therapeutic development. For antibody reagent users, knowing the sequences allows them to perform sequence analysis/alignment to identify binding and cross-reactivity so they can conduct rational experiment design.
Because they share the same mass, isoleucine and leucine are known as isobaric amino acids. Conventional mass spectrometry-based proteomics cannot be easily used to distinguish between isoleucine and leucine.
Amino acid sequencing is the process of identifying the arrangement of amino acids in proteins and peptides. Numerous distinct amino acids have been discovered in nature but all proteins in the human body are comprised of just twenty different types.
Bispecific therapeutics are monoclonal antibodies that carry a specific antigen-binding capability on each arm. Bispecifics are thus capable of having two specificities that can either double the binding affinity of the antibody toward the same antigen (increased avidity), or can now bind to two targets. Bispecifics are most often described as two types: trispecifics and bispecific T-cell engaged antibodies (BiTE).
Written by: María Gerpe, PhD Updated: January 27, 2023 (Published: June 25, 2021) Contents Introduction Types of Antibody Structures Functions of Antibodies Introduction Antibodies or immunoglobulins (Ig) are Y-shaped glycoproteins produced by the adaptive immune system in response to antigens - substances or molecules the immune [...]
Antibodies are integral to life sciences research and therapeutic and diagnostics discovery and development. However, they are inherently prone to variability.
Written by María Gerpe, PhD June 18, 2021 Introduction Research publications represent an additional source of validation proof for commercially available antibodies. As such, academic and industry scientists often also rely on publication references to decide which commercial antibody to purchase. Several independent efforts exist to compile such information. For instance, [...]
Over the past 5 years Rapid Novor has perfected monoclonal antibody sequencing, and is now sequencing mAbs from polyclonal mixtures using REpAb®. After successfully launching their proteogenomics based sequencing technology to deconvolute the immune response, the team has further evolved the technology and has derived the most abundant mAb sequences directly from rabbit blood using only proteomics. The talk will surround the development, progress and use cases for REpAb®.
Over the past several years Rapid Novor has been developing the world’s best antibody protein sequencing platform, sequencing over 2700 monoclonal antibodies and proteins. In 2020, they unveiled their most advanced technology to date - REpAb® polyclonal antibody sequencing. The platform combines the world’s best protein sequencing technology and NGS to comprehensively mine the antigen-specific antibody repertoire present in rabbit and human patient samples. By leveraging the platform, teams can build robust antibody assays and therapeutic leads derived from patients’ blood.
When it comes to polyclonal antibodies, how they are discovered can be just as important as how they are reproduced. In our talk originally presented at Antibody Engineering & Therapeutics Digital 2021, we highlighted the latest technology that’s capable of capturing the most abundant and high-affinity monoclonal antibodies directly from a poly mixture.
At PepTalk 2021, we discussed the importance of antibody standardization and explained why it’s crucial for the longevity of your research. You can listen to the full on-demand video here.
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
If you could have guaranteed stability, certainty, and reproducibility for your research, would you be interested? Imagine this, if you’re 2 years into your project and your freezer died along with all of your important cell lines, what would you do? This is just one of the situations covered in this webinar, along with many other solutions researchers have begun to implement to safeguard their efforts. Whether you’re looking to proceed with stability and certainty or you’re looking for an immediate solution for your current reproducibility challenges, protein sequencing may be the answer.
In this on-demand webinar, our scientific sales executive Jennifer, will briefly cover the fundamentals of protein sequencing, how researchers have benefited from implementing protein sequencing into their pipelines, and discuss how Rapid Novor is able to routinely and robustly achieve 100% accuracy and 100% coverage for both monoclonal and oligoclonal antibodies.
Nowadays, DNA sequencing is so popular that it is easy to forget that the first sequenced biological material was protein – insulin, by Sanger. Sanger, and another researcher, Edman, separately pioneered protein sequencing.
In this on-demand webinar, we discuss why it is important to characterize antibodies based on their physical properties not just by what they bind, and how you can easily do the former via mass spectrometry-based protein sequencing.
One of the most important pieces of information researchers need to know during early stage antibody drug research and development is the sequence information of the antibody protein. With the advancement of mass spectrometry instrumentation and technologies, it is helpful, and sometimes critical, to conduct sequence analysis using mass spectrometry experiments.
Recombinant Monoclonal Antibodies (rAbs) are highly reproducible, customizable and pure alternatives to the traditional antibodies produced by hybridomas. Get the antibody protein sequence, either by DNA sequencing or the de novo protein sequencing technology, you can rest assured that you can have the exact antibody made recombinantly anytime in the future.