Antibody sequencing is a technique that determines the amino acid sequence of an antibody. Discovering the structures and functions of antibodies in living organism is an important tool in understanding cellular processes, which allow drugs that target specific metabolic pathways to be invented more easily.

1. Introduction to Novel
Antibody Sequencing Technique

2. Brief Introduction
Antibody sequencing is a bio-technique that determines the amino
acid sequence of an antibody. Discovering the structures and
functions of antibodies in living organism is an important tool in
understanding cellular processes, which allow drugs that target
specific metabolic pathways to be invented more easily.
This next generation antibody sequencing technique has now
become a routine measurement that can be completed within 10
working days for IgGs. The method is applicable for all antibody
formats, namely IgMs, fluorochrome conjugates, immobilized
antibodies and mixtures. Creative Biolabs, an emerging qualified
service provider in this field, provides researchers their unique
technique in antibody sequencing.

3. With the development of biotechnology, scientists of Creative Biolabs
have developed a novel strategy to determine the complete sequence of
an antibody with unparalleled speed and accuracy. The technique which
combines all the advantages of existing approaches is named “Database
Assisted Shotgun Sequencing” (DASS).
In the first step, the antibody is fragmented to peptides by a special
technique, which generates up to 5000 different peptides per chain.
This set of peptides will be analyzed by high end mass spectrometers to
generate extensive sequence information. MS/MS spectra are then de
novo sequenced by the latest algorithms and matched against a database
with related sequences. Special “in house” data mining tools allow
Creative Biolabs to extract the sequence information from ten thousands
of MS/MS spectra within hours.

4. De Novo Sequencing of the CDR3 Region
While the CDR3 of the light chain is mostly encoded by the
germline sequences, the CDR3 of the heavy chain is usually not
available in databases. It is encoded by the so called D-segments, but
these are modified by nucleases and terminal transferases. Typically,
only 1-4 AA of a D-segment remain in the matured antibody. The
rest of the D-segment is “artificial” and has to go through de novo
sequence. Their method generates many overlapping peptides
during the fragmentation process, enabling them to sequence very
long stretches of unknown amino acids. The high quality of
MS/MS spectra in combination with intelligent data mining, allows
them to read the CDR3 like a book. The technique is so powerful,
that they were able to sequence a 20 kDa protein, which had no
homologue in the database.

5. Sequencing of the V and J and C segments by
"Database Assisted Shotgun Sequencing“
The V and J and C gene segments of antibodies are available in public
databases. During the maturation of an antibody, the B-cell introduces
hypermutations into the sequence to optimize the affinity. Creative
Biolabs’s mapping algorithm is error tolerant and can match the
“mutated” peptides to the corresponding germline reliably.
Because of the great number of peptides, Creative Biolabs is able to get
sequence information for ever bond in the antibody. Typically, 20-70
different MS/MS spectra are generated for each amino acid (AA)
position. Hence, even the hardest sequences of proline and arginine
rich peptides can be resolved.

6. E-mail: marketing@creative-biolabs.com
Tel: 1-631-619-7922
Website: http://www.creative-biolabs.com

7. E-mail: marketing@creative-biolabs.com
Tel: 1-631-619-7922
Website: http://www.creative-biolabs.com