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All You Need to Know Antibody Sequencing
All You Need to Know Antibody Sequencing
Antibody sequencing refers to the definition of the amino acid sequence of an anonymous antibody. The technology of mass spectrometry in de novo antibody protein sequencing has facilitated the performance of antibody sequencing without prior knowledge of DNA sequence or the availability of cell lines.
The protein molecule's mass spectrometry works as a base for the technique of sequencing. It needs 0.2 mg of protein. The period between reporting and analysis has been reduced to a few weeks. The complex protein expression and purification services report contains both light chain and heavy chain sequences, comments on the protein molecule, and the reported sequence's associated data.

Significant Benefits of Antibody De Novo Sequencing 

 

End-to-End Antibody Sequencing

 

Using mass spectrometry techniques with high resolution, optimized settings, including proprietary software for de novo antibody sequencing, means that the entire protein can be sequenced to the c-term from the c-term that includes each amino acid. 

Reproducing an Antibody without a producer cell 

 

If you lose a hybridoma that yields a particular antibody, you can still reproduce the antibody through recombination by only using 0.2 mg of monoclonal antibody protein. Even if you do not possess this much, you may still use other means. 

 

Range of Applications

 

Antibody De Novo Sequencing bears a wide range of applications. The technology can be applied for sequencing all antibody types like IgA, IgG, IgM, Fab, and ScFv, among others, from such animal species as rats, mice, rabbits, and hamsters, if not humans. It has also been applied in sewing conjugated or contaminated antibodies or those with different light chains in pairs. 

 

Unsurpassed Accuracy 

Every amino acid is identified correctly in the sequence since the obtained strong signal signatures in many mass spectrometry peptide readings are available. 

 

Aim of perfection 

The primary purpose of technology is to achieve perfection in mass spectrometric protein sequencing. That has created unending efforts to produce an optimized workflow, including software to enable the sequencing of every protein, however difficult it is, including the maintenance of the highest quality standards. 

 

High Throughput

The mass spectrometry approach for undertaking de novo antibody sequencing is essential, with one or hundreds of antibody molecules awaiting sequencing. 

 

Reportage of Everything 

 

Using antibody de novo sequencing has often facilitated the detection of differences that are not expected in novel glycosylation sites or sequencing not detected by DNA sequencing.

 

Databases of Antibody Sequence 

 
 

UniProt  

UniProt refers to a high-quality, comprehensive, and freely accessible resource of functional information and protein sequence. It has no specific focus on antibodies but only includes representative germline antibody sequences set with a limited population of non-germline antibody sequences. 

 

IMGT 

This refers to the international ImMunoGeneTics information system that serves as a global reference for the high-quality integrated knowledge resource specializing in immunoglobulins (Ig), T cell Receptors, and other Major Histocompatibility Complex molecules of vertebrates, including humans. IMGT composes seven databases that encompass antibody genome, sequence, and structure. For instance, the monoclonal antibodies IMGT database called IMGT/mAbDB offers an antibodies' expertise resource on monoclonal antibodies or antibodies with clinical indications. 

 

ABCD database (AntiBodies Chemically Defined database)

 

The ABCD Database was launched in 2020 as a sequenced antibodies repository, developing integration of curated information on the antibody, including its antigen, with cross-links to protein and chemical entities databases. Every antibody is provided with a unique identification number used in academic publications for increasing experiments' reproducibility. 

 

Thera-SAbDad 

This is also called Therapeutic Structural Antibody Database. It tracks all therapeutics related to antibodies- and nanobodies recognized by the World Health Organization (WHO) and identifies any SAdDab corresponding structures with exact or near-exact sequence matches of variable domains. 

 

 

Final Thoughts