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Polyclonal Antibody Sequencing

Polyclonal Antibody Sequencing Directly from Protein

Polyclonal antibodies are commonly used reagents for diagnostics and research assays.  They are derived from the serum of immunized animals, and their exact composition is not easily determined.  There are many benefits to converting your polyclonal antibody into recombinant monoclonals.

Product Consistency

Eliminate batch-to-batch variability of your antibody and create an infinite supply.

Intellectual Property

Protect your IP by protecting the component sequences of your antibody.

Flexibility

Engineer, conjugate, and reformat your product into new tools with efficiency.

How does polyclonal antibody sequencing work?

Stage 1: Clonality Assessment

A  shallow first-past assessment of sequence diversity, particularly in the CDR3, reveals the level of complexity of the sample.  Too many CDR3s indicates that only the most abundant will be sequenceable.  Simplification of the sample can be a good option to fractionate the polyclonal antibody into desired reactivity groups.

The CDR3 network to the right shows two major families of CDR3s present in a polyclonal antibody.  Each node is a CDR3 sequence, and edges are created between CDR3s that have similar sequences.

Polyclonal antibody sequencing stage 1 sequences CDR3s and clusters similar antibodies to assess clonality.

Stage 2: Assembly and Polyclonal Antibody Sequencing

A deep second-pass analysis generates candidate sequences.  Each sequence is an assembly of peptides, and the sequences are reviewed in silico to eliminate unnatural sequences and structures.  The figure shows the peptide support for one candidate VHH sequence.

Many peptides are assembled to perform polyclonal antibody sequencing.  The image shows the map of peptides to a final sequence.

Stage 3: Expression and Validation

The final stage of polyclonal antibody sequencing is recombinant production and testing.  CDR3 clustering groups candidate sequences into families.  Representatives from each family are selected for recombinant expression and binding validation to the target.

Additional in silico screening for stability can be applied to remove sequences that are unlikely to fold correctly and bind the target.

In silico analysis can predict stability before spending time and effort on expression and testing in polyclonal antibody sequencing.

Have access to more than just polyclonal antibody?

Alicanto is a sensitive, accurate technology that uses both B cells and serum for polyclonal antibody sequencing and antibody discovery.

If you have B cells, PBMCs, or access to the host animal, Alicanto can be a great option for polyclonal antibody sequencing.

Looking to sequence monoclonal antibodies or mixes of monoclonals?

Our Valens technology is best suited for sequencing monoclonal antibodies or low complexity mixes of antibodies.

Based on almost 20 years of technology development, Valens is a highly accurate, fast technology for sequencing antibodies.

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