Creative Biolabs provides an innovative de novo antibody sequencing service platform based on the state-of-the-art high-resolution mass spectrometer, with DASS (Database Assisted Shotgun Sequencing) technology and rich bioinformatics analysis experience, allowing for fast and accurate analysis of antibody primary amino acid sequence information.
De Novo Antibody Sequencing Workflow
Creative Biolabs' de novo antibody sequencing services require a minimum of 50 µg of purified antibody to achieve 100% coverage and unparalleled accuracy. Our service workflow adheres to the following streamlined process:
- Using several trypsins to initially dissolve the antibody samples into overlapping peptides.
- Optimization of fragmentation after LC-MS/MS digestion.
- Characterizing the overlapped peptides by de novo peptide sequencing.
- Automated assembly of the antibody sequence using the characterized peptides.
- Detailed data analysis using state-of-the-art bioinformatics methods to accurately determine the amino acid sequence of the digested peptide.
Intact mass validation of the final antibody sequence.
Aiming to Rescue Valuable Samples with Minimal Consumption
- From a small portion of your valuable sample, we have a possibility of obtaining the sequence of more than 90%.
- Even extremely diluted samples (e.g. western blot antibody) can be enriched for sequencing.
- Some matrix-coupled mAbs are also compatible with our sequencing techniques, but additional samples could be required.
- The kappa chain of myeloma MOPC 21 is one of the two light chains that many older hybridomas can produce. Even when the MOPC 21 kappa chain is two times molar excess in these mixes, sequencing is typically still possible.
- In some cases, if there is BSA in the sample, we can remove it. We can also sequence mouse IgG in hybridoma supernatant that contains bovine serum antibody.
- We also offer DASS-based sequencing services for proteins and enzymes, including full end-to-end sequencing, determination of post-translational modifications, intact mass, N-glycosylation sites, artificial tags, etc.
Support for multiple species antibodies
Our method works with all antibody formats from all species (mouse, rat, chicken, hamster, llama, rabbit, etc.; Fab, scFv, etc.).
Human/Humanized Antibody Sequencing
Human/humanized antibodies are the growing trend in the global monoclonal antibody market.
Access to precise antibody sequence information, facilitating the discovery of humanized/humanized therapeutic antibodies.
Rodent Antibody Sequencing
Hybridoma technology is used for various rodent antibody production, and applied for relevant model animal-based research.
Highly precise and successful sequencing for dozens of rat, mouse, and hamster antibodies.
Rabbit Antibody Sequencing
Rabbit antibodies with higher dilution ratios and better labeling efficiency are an exceptional source for antibody applications.
Capable of overcoming the sequencing challenges of rabbit antibodies as carrying multiple glycans in the heavy chain Fc region and an additional SH bridge.
Chicken IgYs Sequencing
Chicken IgYs are ideal immunological detection reagents given their higher affinity with persistent titers and scalable production.
Determination of the actual sequence of IgYs favors their biomedical and biochemical applications.
Support for multiple isotypes
IgG antibody sequencing. IgG is the major antibody component of serum and plays a role in the immune response by activating complement and neutralizing multiple toxins for antimicrobial and antiviral purposes. IgG antibody sequencing provides insights into their diversity, contributing to vaccine development, therapeutic antibody engineering, and understanding immune responses.
IgM antibody sequencing. IgM is mainly found in blood vessels and has strong cytotoxic and cytolytic activity. It is the primary antibody against intravascular infections, which plays a crucial role in recognizing and neutralizing pathogens. IgM antibody sequencing provides a detailed understanding of their composition, aiding researchers in unraveling immune responses and designing targeted therapies. As the constant region of IgM is modified by several glycans, which cannot be cleaved by PNGase F, we can only guarantee complete coverage for the variable part of the antibody, and slightly more samples may be required. Case Study for De Novo Sequencing of an IgM Antibody
IgA antibody sequencing. IgA is the primary antibody for localized anti-infection immunity of the body's mucous membranes, targeting pathogens such as bacteria and viruses on mucous membrane surfaces. Sequencing of IgA antibodies helps to provide insights into the immune system's response to infections in mucous membrane sites, such as the respiratory tract, as well as to help in the identification and characterization of new pathogens.
IgE Antibody Sequencing. IgE antibody triggers an allergic reaction by binding mainly to allergens. Sequencing of IgE antibodies allows for accurate diagnosis and treatment of allergic diseases and determines how the body reacts to certain allergens.
Coupled Antibody Sequencing. Especially for coupled antibodies (e.g., APC, PE, Fluorescent dyes, FITC coupled antibodies), de novo antibody sequencing at Creative Biolabs is realized to remove the interference of precise sequencing caused by the conventional methods of existing covalent binding of dyes. Larger protein ligands (PE, APC) make sequencing slightly more challenging. However, this can be compensated by using a higher protein concentration, because sequencing quality is dependent on sample amount.
Single Domain Antibody (sdAb) sequencing. sdAb features both the high specificity of traditional antibodies and the excellent penetration of small molecule drugs, delivering high potency in downstream engineering. Sequencing of soluble and functional sdAb from diverse sources can be achieved by subunit and is also applicable to sdAb conjugates or immobilized sdAbs.
We have over 90% chance to get the sequence from a tiny amount of your precious sample (e.g. 20 µg) and can enrich even a very dilute sample (e.g. western blot antibody) for sequencing. We also provide DASS-based Protein and Enzyme Sequencing services including full end-to-end sequencing, determination of post-translational modifications, intact mass, N-glycosylation sites, artificial tags, etc.
Our De Novo Antibody Sequencing Advantages
Precise discrimination of Leucine & Isoleucine by mass spectrometry.
98% of the sequenced mAbs show equal or better performance compared to the originator.
We promise to completely cover the VJ and VDJ regions. Otherwise, you will not be charged.
We have sequenced 1000+ mAbs yet.
Sequence antibody conjugates, BSA containing mAbs lama or rabbit mAbs, IgMs, or even immobilized mAbs.
Get full coverage from 50-100 µg of protein.
Our method is high throughput compatible. Up to 200 mAbs/year, we would be happy to arrange your HT project.
An expert R&D team committed to protocol optimization to offer high-quality data.
Packages Available
| Choose Your Package | Basic DASS Package | Advanced DASS Package |
|---|---|---|
| Small sample quantity required | 50-100 μg | 100 μg |
| Applicable to all antibody types and species | √ | √ |
| 100% coverage of VJC and VDC sequence | √ | √ |
| Leu/Ile discrimination method | Chymotrypsin digestion and codon analysis | Mass spectrometry |
| Accuracy of Leu/Ile positions | Inferred Leu/Ile positions | Precise Leu/Ile determination |
| CDR3 Region Sequencing | √ | √ |
| MS/MS data and spectra provided on request | √ | √ |
| Timeline | 10 working days | 2~4 weeks |
Our Guarantee
Full Coverage
100% coverage of the VJ and VDJ region.
Multiple Cleavages
At least 4 different peptides will cover each amino acid position.
Sequence Validation
The determined sequences should match with the measured intact masses of heavy (± 1.8 Da) and light chain (± 1.2 Da).
Fast Delivery
Get your sequencing within 10 working days. 2~3 additional weeks are needed if the Ile/Leu discrimination is required.
$18,000 for sequencing a package of 2 antibodies!
Sample requirements:
- Both samples are sent simultaneously.
- Monoclonal antibodies from mouse, rat, rabbit, hamster, camelidae, or other species.
- IgG, Fab, scFv, VHH, or other formats.
- 50-100 µg with or without BSA/HAS.
Numerous successful cases from Creative Biolabs have demonstrated that we are qualified to provide 100% accurate antibody sequencing services with a satisfaction guarantee. Please inquire us for a discussion about your project needs.
