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: 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: IVD and immunoassays, supply chain risk management considerations How monoclonal and polyclonal antibody sequencing works, requirements Effective production, validation, and scale-up of recombinant antibody reagents Characterization of mAbs for assay design Abstract Polyclonal antibodies (pAbs) are well known for their robustness and [...]
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: Vanessa Yoon Calvelo, PhD Published: December 22, 2022 Contents The Great Debate in Antibody Discovery and Generation Traditional Antibody Discovery and Generation Technologies Emerging Antibody Discovery and Generation Technologies Antibody Discovery and Generation Technologies – Advantages and Disadvantages World’s First: Decoding Serum pAbs with Rapid Novor’s REpAb® Technology [...]
Written by: Vanessa Yoon Calvelo, PhD Updated: January 18, 2023 (Published: September 15, 2022) Contents Analyzing Biomolecular Interactions is Fundamental to Understanding Biological Systems Analytical Techniques for Biomolecular Interactions SPR Applications for Biomolecular Interactions SPR – A golden standard for biomolecular interaction analysis Analyzing Biomolecular Interactions is [...]
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 two [...]
Written by: Vanessa Yoon Calvelo, PhD Published: July 21, 2022 Contents Mass spectrometry-based approaches for structural analysis of proteins HDX-MS Applications for ‘Difficult’ Proteins HDX-MS – A powerful analytical technique for protein characterization Mass spectrometry-based approaches for structural analysis of proteins Characterization of proteins and protein [...]
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 Characterization of PTMs by next generation protein sequencing The Importance of Post-Translational Modifications (PTMs) Post-translational [...]
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 [...]
Written by Yuning Wang, PhD May 31, 2022 Contents Background What is Polyclonal Antibody Sequencing? Applications of Polyclonal Antibody Sequencing Advantages of Polyclonal Antibody Sequencing World’s First: De Novo Polyclonal Antibody Sequencing at Rapid Novor Background The native immune system produces polyclonal antibodies (pAb) by different B-cell [...]
Written by Yuning Wang, PhD May 13, 2022 Contents What is HDX-MS? HDX-MS Workflow Applications of HDX-MS HDX-MS Advantages HDX-MS Epitope Mapping of Antibodies HDX-MS Epitope Mapping at Rapid Novor What is HDX-MS? The origin of hydrogen-deuterium exchange (HDX) dates back to the 1950s, when protein scientist [...]
Written by: Yuning Wang, PhD Updated: January 26, 2023 (Published: March 15, 2022) Contents What are CDRs? Antibody CDRs De Novo Protein Sequencing as a Tool for Identifying CDRs Sequences Applying De Novo Protein Sequencing to Identify CDRs Sequences Annotation Schemes for Identifying CDRs by Sequence Rapid Novor can Help with [...]
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 [...]
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 María Gerpe, PhD January 2, 2022 Contents What are Recombinant Antibodies? What is the Difference Between Recombinant and Traditional Antibodies? Why are Recombinant Antibodies Important? Types of Recombinant Antibodies How are Recombinant Sequences Obtained? Additional Resources References What are Recombinant Antibodies? [...]
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.
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
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.
Written by Yuning Wang, PhD August 1, 2021 What is DNA Sequencing? 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 [...]
Written by: María Gerpe, PhD Updated: January 26, 2023 (Published: July 23, 2021) Contents Why is it Important to Know the Protein Sequence? MATCHmAbTM Confirms the Primary Amino Acid Sequence Critical to Both Specificity & Reproducibility Why is it Important to Know the Protein Sequence? As proteins [...]
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
Proteins are composed of peptide chains, which in turn are made up of a string or linear sequence of amino acids (Figure 1A). 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 (Figure 1B). As such, the primary structure of a protein is typically recorded starting at the amino-terminal (N) end and continuing to the carboxyl-terminal (C) end. The primary protein structure may be directly sequenced from a sample of the protein itself or inferred from the DNA sequence.
Protein mass spectrometry refers to the use of mass spectrometry in the study and characterization of proteins, including their quantification, profiling, interaction mapping, and identification of their post-translational modifications (1,2). Protein mass spectrometry may also be referred to as mass spectrometry-based proteomics. Mass spectrometry-based proteomics consist of three approaches: top-down, middle-down, and bottom-up proteomics
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
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 [...]
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, [...]
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
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
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
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
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.
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
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.