Written by María Gerpe, PhD 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? Recombinant antibodies are [...]
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
In-Depth Characterization of Monoclonal Antibodies with a Single Experiment and Fully Automated Data Analysis
To develop robust mAb biologics, it is vital to fully characterize the protein, including its primary sequence, mutations, and important post-translational modifications
In this study, we conducted a large-scale statistical analysis of protein sequencing data from samples digested with multiple proteases to understand the impact of using different combinations of proteases to improve the depth of sequence coverage in the application of de novo protein sequencing.
Anti-drug antibody (ADA) assays are critical to assess the clinical efficacy and safety of a biological drug and rely on control reagents that mimic the ADA response to the biological drug being tested. These positive controls typically consist of animal-derived pooled polyclonal antibodies or human monoclonal antibody reference panels against the target protein drug.
Abstract In this on-demand webinar, we 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. Webinar details [...]
Abstract 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. Key Takeaways Antibodies are generally characterized based on what they bind, not their [...]
Overcoming Irreproducibility in Life Science Research Sponsored by: Rapid Novor, Inc & Absolute Antibody 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. Full Webinar Panelists: Andrew Bradbury, CSO at Specifica “Drug-Like Antibodies from [...]
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.
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.
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 July 23, 2021 Why the Amino Acid Sequence Matters 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 therefore function. The importance [...]
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
The protein sequence is key to understanding the function of a protein target and is critical to therapeutic and diagnostic development. This is particularly important for antibodies whose code diversity and glycosylation impact both function, and stability.
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).
Antibodies are integral to life sciences research and therapeutic and diagnostics discovery and development. However, they are inherently prone to variability.
Monoclonal antibodies (mAbs) are widely used in research, diagnosis, and pharmaceutical purposes. Lately, the relatively lower quality of research-purpose mAbs is a point of concern within the research community.
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, [...]
A Large-Scale Comparison of MS-based Antibody De Novo Protein Sequencing and Targeted DNA Sequencing
The DNA sequences of antibodies are highly diverse due to the V-(D)-J recombination and hypersomatic mutations. As such, relying on homology-based searches to sequence novel antibodies can introduce bias to sequences obtained from proteomics approaches.
Mouse monoclonal antibodies (mAbs) are highly attractive for manipulation for therapeutic applications as their manufacturing is relatively easy and well-established compared to mAbs derived from larger animal models. However, they also pose several challenges which limit their use as therapeutic agents.
In-vitro diagnostics (IVDs) are one of the most commonly used tools to diagnose conditions and guide treatment decisions and are often considered the “silent champion” of healthcare. They work by detecting the absence or presence of particular markers or by measuring the concentration of analytes or specific substances.
Research Insurance with de novo Protein Sequencing Originally presented October 16, 2020 by Monique Seymour, International Scientific Sales Executive, Rapid Novor 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 [...]
We are able to sequence any species because we use de novo protein sequencing by tandem mass spectrometry (MS/MS). This type of protein sequencing by mass spectrometry does not rely on existing databases that might introduce bias or wrong amino acid calls.
We recommend that you ship your samples either lyophilized or in solution. If your samples are already lyophilized it is best to ship them to us in that form, they can be shipped at ambient temperatures, and are suitable for transfers longer than 24h.
How much protein and what purity is needed for protein sequencing? A: We typically accept samples at 100 µg with at least 80% purity, rare in the field. To measure the amount of protein, you can use a microvolume spectrophotometer such as the Nanodrop or a traditional Bradford assay.
The most common protein contaminants include keratin and serum albumin. Other Mass Spectrometry (MS) protein contaminants identified in research labs worldwide by a recent Human Proteome Organization study are casein and E. coli proteins.
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.
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.