July 2022

Protein Characterization by HDX-MS

2023-06-21T11:01:38-04:00July 26th, 2022|Articles|

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.

June 2022

Why do Post-Translational Modifications Matter?

2023-05-15T11:19:45-04:00June 14th, 2022|Articles|

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 [...]

Protein Structure and How to Study It

2023-05-15T11:21:16-04:00June 10th, 2022|Articles|

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 [...]

May 2022

What is HDX-MS Epitope Mapping?

2023-06-21T11:00:11-04:00May 15th, 2022|Articles|

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.

November 2021

September 2021

Structure of an Amino Acid

2024-05-06T12:51:35-04:00September 14th, 2021|Articles|

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.

Why is Protein Sequencing Useful?

2024-05-06T12:53:45-04:00September 3rd, 2021|Articles|

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.

August 2021

What are Monoclonal Antibodies?

2023-05-15T11:39:30-04:00August 25th, 2021|Articles|

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

How to Determine Peptide Sequences

2023-05-15T11:41:09-04:00August 17th, 2021|Articles|

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.

Key Pain Points in Amino Acid Sequencing & How to Avoid Them

2023-05-15T11:42:16-04:00August 13th, 2021|Articles|

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.

Antibody Validation and its Use Cases

2024-04-29T13:22:17-04:00August 4th, 2021|Articles|

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.

De Novo Protein Sequencing vs DNA Sequencing

2024-04-01T15:52:10-04:00August 1st, 2021|Articles|

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.

July 2021

Why Verifying the Protein Sequence Is Crucial for Antibody Validation

2023-06-21T10:46:18-04:00July 23rd, 2021|Articles|

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.

What is the De Novo Amino Acid Sequencing Method?

2024-04-11T11:50:52-04:00July 19th, 2021|Articles|

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.

What is Protein Mass Spectrometry?

2023-06-21T10:44:34-04:00July 18th, 2021|Articles|

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.

How Do You Obtain the Sequence of an Antibody?

2023-05-15T11:49:55-04:00July 10th, 2021|Articles|

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.

Isoleucine and Leucine

2023-05-15T11:51:04-04:00July 7th, 2021|Articles|

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.

What is Amino Acid Sequencing?

2023-05-15T11:52:07-04:00July 5th, 2021|Articles|

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.

June 2021

March 2021

May 2020

Introduction to De Novo Protein Sequencing

2023-06-21T10:54:37-04:00May 21st, 2020|Webinars|

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.

August 2019

April 2019

February 2017

Antibody Protein Sequence Analysis Using Mass Spectrometry

2023-05-15T11:58:18-04:00February 3rd, 2017|Articles|

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.