Antibody Sequencing Service

Service Description

Getting accurate sequence of the variable domains of your monoclonal antibodies will open up a realm of possibilities for downstream antibody engineering from in vitro recombinant expression to affinity maturation and humanization. Knowing the sequence of your antibodies will protect you from the loss of your valuable clones; it is one of the first steps to get protection of your antibodies through patent applications and it is an important quality control to maintain consistent cell lines for batch production.

  • Clone protection.

  • Patent applications & Intellectual property.

  • Recombinant antibody expression.

  • Antibody engineering & Drug development.

  • Stable cell line quality control & Traceability.

Antibody Design Labs offers a fast, reliable and competitively-priced sequencing service to clone your antibody cell line. Both light chain and heavy chain variable domain cDNAs will be amplified using either a combination of leader primers or state-of-the-art rapid amplification of DNA ends (5’-RACE). A full length sequencing service including complete leader sequence using 5’-RACE and sequence of constant domains (full antibody) is also available upon request. We can sequence from a variety of species including mouse, rat, rabbit and hamster hybridomas or human EBV-immortalized B-cell lines and isotypes including IgM & IgA. Hybridomas are prone to contain multiple chains as a result of cellular fusions involving multiple B-cells (quadromas), non-productive rearrangements generated during B-cell maturation and pseudogenes; we will conduct a thorough and extended analysis of multiple clones to confirm the identity of the relevant full-length, in-frame sequences and eliminate aberrant endogenous myeloma light chains. This is a “No Success – No Fee” service; you pay on delivery of your antibody sequence.

General Procedure

  • Total RNA extraction and cDNA Synthesis.

  • 5’RACE extension.

  • Amplification of VH and VL domains including leader sequence and partial constant regions CH1 and CL.

  • Cloning of PCR positive reactions.

  • Colony PCR and sequencing of clones with proper insert size.

  • Sequencing analysis up to 5x coverage or 10 clones per chain, whichever come first.

Sequencing Services

  • V Domain Sequencing Service. Identify variable domains using primer sets, perfect for routine work during antibody generation and re-cloning as scFv. Mutations, normally silent, may be introduced at junctions; not all species available, please inquire. Delivery Time: 2 weeks – Price: starting from $950.
  • V Domain Sequencing plus Leader. Identify variable domains and leader sequences using 5’RACE reactions; this is the standard way in the field to assert the exact sequence of the variable domains of your antibody and important step to take when translating an antibody into clinic. Delivery Time: 3 weeks – Price: starting from $1450.
  • Antibody Sequencing plus Test Expression. Identify variable domains and leader sequences using 5’RACE, clone and express the identified antibody in 10 ml culture for testing. Delivery Time: 4-6 weeks – Price: starting from $2900
  • Full-Length Sequencing Service. Identify leaders, variable domains and constant regions, important to verify cell lines and identify phenotypic variants. Delivery Time: 4 weeks – Price: starting from $2250

Pricing given for mouse & rat hybridomas; charges apply for hamster, rabbit, primate & human cell lines.

Project Management

Sample Submission

A detailed submission protocol will be provided. Information on species and isotype will greatly accelerate turnaround time. You need to ship between 1 and 5×10(6) cells per hybridoma either as a flash-frozen cell pellet on dry ice or as RNA-stabilized cells at room temperature (we provide containers with RNA stabilization reagent upon request). We accept smaller samples 1×10(5) and less and can help you to recover the sequence of dead hybridomas.

Deliverables

  • Detailed project report.

  • Annotated sequences including CDR identification & sequencing traces.

  • Plasmid map of each clone.

  • Spin-column purified plasmid DNA for each clone with a certificate of analysis and package tracking (shipping charges apply).

We welcome discussing the result directly with you.

Additional Services

  • Isotyping service to establish the exact isotype.

  • Cloning & expression of recombinant scFv or immunoglobulin.

  • Binding analysis by ELISA to confirm that the variable domain sequences have been properly identified.

Cost & Turnaround Time

See table above for starting service fees & expected turnover.

Request Service

P

lease, request a specific quote by completing the online quotation form accessible by pressing the following button; you will be contacted back by the next business day.

Request A Quote

For all other inquiries, contact us by email at info@abdesignlabs.com or by phone at 1-877-223-3104 (PST). We guarantee competitive pricing.

All our work is performed by highly experienced staff at our San Diego facility in California. We use highly efficient cloning techniques to ensure timely delivery and have very high standards for validation. All products and services are for research use only and are not intended for use in humans.

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