Disulfide Bond Analysis.
Identification and quantification of disulfide bonds by LC-MS, designed for monoclonal antibodies (mAbs).
How Disulfide Bond Analysis Works.
Disulfide bonds play an important role in stabilizing the spatial structure of proteins, maintaining correct folding conformation, maintaining and regulating their biological activities. The formation of disulfide bonds forces the amino acid residues in different regions of the same or different peptide chains to converge together, so that the peptide chains quickly fold and form a stable spatial topology.
HPLC-MS/MS is a powerful tool for protein characterization due to its high sensitivity, high accuracy and abundant structural information. The common method of disulfide bond analysis by mass spectrometry is analyzing the protein digest mixtures after enzymatic digestion, then identifying the type and location of disulfide bonds. We typically use trypsin and chymotrypsin digested non-reduced samples, which are then separated by Liquid Chromatography and analyzed by tandem mass spectrometry (MS/MS).
Why Disulfide Bond Analysis Matters.
Disulfide bonds form between pairs of cysteines and they join the parts of an antibody together. In antibody engineering, a modified antibody sequence may not have the desired disulfide bonds present. The bond may form between the wrong cysteines some of the time. Discovery scientists and protein engineers may analyze disulfide bond shuffling in order to select or reject mAbs for downstream work. Production teams who have access to this information early in their process can have higher success rates downstream.
Getting Started.
Requirements
- Amino acid sequence(s) in FASTA format
- 20 μg per analysis per sample
- >90% purity
- Monoclonal antibody (any isotype, format or fragment)
Deliverables
- Sequence annotated with disulfide bonds
- Confidence measure on each disulfide bond assignment
- Prevalence of each disulfide bond configuration (percentage)
- Optional: comparison of samples, if multiple samples are given
Timeline
Screening
Talk to Our Scientists.
We Have Sequenced 9000+ Antibodies and We Are Eager to Help You.
Through next generation protein sequencing, Rapid Novor enables reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and developed the first recombinant polyclonal antibody diagnostics.
Talk to Our Scientists.
We Have Sequenced 9000+ Antibodies and We Are Eager to Help You.
Through next generation protein sequencing, Rapid Novor enables timely and reliable discovery and development of novel reagents, diagnostics, and therapeutics. Thanks to our Next Generation Protein Sequencing and antibody discovery services, researchers have furthered thousands of projects, patented antibody therapeutics, and ran the first recombinant polyclonal antibody diagnostics
Talk to our scientists. We have sequenced over 9000+ antibodies and we are eager to help you.
Other Antibody Characterization Services.
Rapid PTM Analysis
Locate, identify and quantify post-translational modifications. Compare between samples.
Sequence Variant Assessment
Identify and quantify variants of your intended antibody within your sample, and consider other contaminants.
Glycosylation Analysis
Find glycosylation site occupancy, and profile their relative abundance %. Compare many samples.
Peptide Mapping
Maximize coverage and confidence, map, identify and quantify post-translational modifications. Compare between samples.
Disulfide Bond Analyses
Analyse peptides cross linked by disulfide bonds. Quantify disulfide bond shuffling in engineered constructs.
Intact Mass, Custom Analyses
Please inquire about intact mass analysis, subunit analysis, etc.
Other Services.
Antibody Sequencing & Discovery Services.
REmAb®
Monoclonal antibody sequencing from small antibodysamples, no need for hybridoma or DNA information. Full sequence in record time.
Explore REmAb
REpAb®
Polyclonal antibody sequencing and antibody discovery service. Sequence antibodies from blood or a polyclonal mixture.
Explore REpAb
MATCHmAb™
Antibody verification by real-time peptide mapping. Antibody sequence confirmation service for reproducibility.
Explore MATCHmAb
HDX-MS
Mass spec based epitope mapping. Epitope mapping for identify the binding site of an antibody to its corresponding antigen with the highest confidence and resolution.
Explore Epitope Mapping
SPR Analysis
Label-free evaluation of antibody-antigen binding relies on a Surface Plasmon Resonance technology. Combined with our expertise in antibody interactions, this results in high sensitivity and the lowest noise even in complex analytes.
Explore SPR Analysis
NovorIg™
Quantify the immune response by accurately profiling the vast antibody-protein repertoire and its different isotypes, and subtypes in serum over time.
Explore Immune Response Profiling