Antibody Sequencing Service

Service Details

Note:
a) Sample request:
Cell types: Hybridoma cells, B cells, Single B cells
Amount: >1×10⁶ cells
Species: mouse, rat, rabbit, ferret, etc.
b) Additional services: High-throughput recombinant antibody expression, Hybridoma antibody production, Antigen-antibody binding confirmation

Case Studies

• Case 1: Typical sequencing report for Sanger
  

  
• Case 2: High-throughput NGS antibody sequencing for 4-1BB antibody development

  Goal: Develop antibodies specific to 4-1BB

  Immunization: 4-1BB extracellular region

  Result: 107 4-1BB antibodies were obtained and the genome information was analyzed by the high-throughput NGS antibody sequencing platform

  1. Phylogenetic Tree Analysis - VH Region

  

  Out of 107 clones, 5 clones show the identical V-region with other clones

  2. Phylogenetic Tree Analysis - VH Region

  

  Out of 107 clones, 6 clones show the identical V-region with other clones

  3. Germline Analysis (Indicates antibody diversity)

  

  

  4. CDR Length Analysis (Indicates antibody candidate)

  

  

Additional Services

FAQs

• What is the principle of hybridoma cell sequencing?

  Hybridoma sequencing involves amplifying and sequencing the cDNA encoding the VH and VL domains from a hybridoma cell line. By utilizing high-throughput sequencing technologies, this technique enables the rapid and efficient determination of antibody gene sequences.

• What are the advantages of hybridoma cell sequencing?

  1. Preserved Antibody Diversity: Hybridoma cell sequencing allows for the sequencing of antibodies produced by hybridoma cells, which are derived from a specific B cell clone. This preserves the natural diversity present in the immune response. In contrast, de novo antibody sequencing relies on isolating individual antibodies and may not capture the full diversity present in the immune response.

  2. Time and Cost Efficiency: Hybridoma cell sequencing can be a more time and cost-efficient approach compared to de novo antibody sequencing.

  3. Quality Control and Validation: Since the sequences are obtained from the hybridoma cells themselves, it ensures that the sequenced antibodies are indeed the ones produced by the hybridoma clone of interest. In de novo sequencing, additional validation steps are required to confirm the authenticity of the obtained sequences.

  4. Compatibility with Established Hybridoma Lines: The hybridoma cell lines may have a known history, desired properties, and a documented track record of antibody production. Sequencing the antibody genes from these hybridomas provides a wealth of information about their antibodies, aiding in further characterization and development.

  5. Distinguishing certain amino acids: Protein sequencing can encounter challenges in differentiating specific amino acids, such as leucine and isoleucine, which may introduce errors in the final sequence. In contrast, hybridoma sequencing directly sequences the cDNA of the antibody-producing cells, eliminating this potential issue and yielding a more precise, accurate, and reliable result.

• How to choose the antibody sequencing method, Sanger or NGS?

  1. Research Objectives: If you require the full-length coding sequence of the antibody, including the variable region and constant region, Sanger sequencing may be a suitable choice. However, if you need a comprehensive analysis of the antibody repertoire, including clonal diversity and identification of rare or low-abundance variants, NGS can provide a more comprehensive picture.

  2. Sample Quantity: Assess the quantity of samples you need to sequence. Sanger sequencing is generally suitable for a smaller number of samples due to its labor-intensive and costly nature. On the other hand, NGS platforms offer higher throughput, enabling the sequencing of a larger number of samples simultaneously.

  3. Cost Considerations: NGS is generally more cost-effective than Sanger, especially for a large number of samples. Sanger sequencing is more complex and typically has a higher cost. However, if you require full-length coding sequence of the antibody, Sanger sequencing remains the more suitable choice.

  4. Sequence Accuracy: Assess the level of sequence accuracy required for your research. Sanger sequencing has a long-standing reputation for high accuracy, particularly in obtaining individual full-length sequences. NGS platforms, while generally accurate, may have higher error rates, especially in homopolymeric regions or repetitive sequences. Consider whether the specific accuracy requirements of your project can be met by either method.

Quotation and Ordering

Our customer service representatives are available 24 hours Monday through Friday. You may contact us anytime for assistance. Orders can be placed online, over the phone, by email or fax. Please have your payment information (credit card, purchase order or bank information) ready.