Hybridoma/Monoclonal Antibody Sequencing

Monoclonal antibody sequencing includes hybridoma cell line sequencing and clonal B cell sequencing. Both of these kinds of cells can produce a single antibody against a unique antigen after screening. Synbio Technologies uses 5’RACE and Sanger sequencing technologies to provide both accurate and efficient hybridoma/clonal B cell sequencing services across a variety of species.

standard-hybridoma-sequencing-process

Standard Hybridoma Sequencing Process

Standard antibody variant region PCR amplification primers design schematic

hybridoma-sequencing

mRNA isolated from hybridoma samples is subjected to RT-PCR with unique species-specific primer sets to amplify the target regions. Synbio Technologies’s antibody variable region sequencing service has identified more than 10,000 hybridoma cell lines. More than 95% of the samples have been successfully sequenced with both the heavy chain and light chain identified. Even with technically difficult samples, Synbio Technologies’s approach has demonstrated a high success rate in numerous cases. Synbio Technologies’s service, coupled with online data tracking and management system, provides customers with absolute confidence at every step.

Following the successful cloning of variable regions, the sequences can be fused to a variety of IgG or IgM backbones. Alternatively, variable region sequences can be reformatted as scFv or bispecific antibodies in various configurations. Transiently produced antibodies can then be validated and characterized by downstream analytical services.

Competitive Advantages

  • Comprehensive: Sequencing of heavy and light variable regions or full-length of antibody sequences.
  • Wide applicability: Relying on a professional immunogenetics database and our rich experience in antibody sequence analysis and design, antibodies from any species (mouse, rat, human, rabbit, hamster, camel, sheep, etc.) and various antibody subtypes (IgG1, IgG2a, IgG2b, IgG3, IgG4, IgM, IgE, IgA etc.) can be designed and sequenced.
  • Reliable: At least 4 positive clones are selected for Sanger sequencing and sequence analysis is performed to exclude any invalid sequences.
  • High-throughput and Cost-Effective: Whether it is a sample or hundreds of samples, we can do it all. The more samples that are provided, the lesser the cost.
  • Fast turnaround time: We provide express sequencing services, as fast as one week.
  • One-stop service: We provide a one-stop solution from hybridoma sequencing to recombinant antibody production. For the non-human antibodies, our antibody humanization service can be considered.

Cell Sample Preparation and Shipping

  • For hybridoma cell lines or clonal B cells: customers will need to submit hybridoma cell samples (>1×105 cells) and indicate the species and subtype of the monoclonal antibody.
  • There are two types about the shipping:
    I: fresh cells in frozen vial(s) and shipped in dry ice.
    II: 2. wash cells with PBS and re-suspend the cells in 5–10 volumes (~1ml) RNAlater Solution, then ship with blue ice.

Services Specifications

Service Cat#DescriptionTurnaround TimePrice
AHS001Cloning and sequencing of mouse hybridoma IgG/IgM variable regions1-2 WeeksStarting at $800
AHS002Cloning and sequencing of rat hybridoma IgG/IgM variable regions1-2 WeeksStarting at $800
AHS003Cloning and sequencing of hamster hybridoma IgG variable regions1-2 WeeksStarting at $800
AHS004Cloning and sequencing of rabbit hybridoma IgG variable regions1-2 WeeksStarting at $1500
AHS005Cloning and sequencing of human hybridoma IgG variable regions1-2 WeeksStarting at $1500
AHS006Cloning and sequencing of other species hybridoma IgG variable regions2-4WeeksStarting at $2000

Deliverables

  • Raw sequence data (100% accuracy guarantee, otherwise there is no fee)
  • Final sequence report with variable heavy chain and light chain sequences or full length sequence and specific regions annotated
  • Plasmid contains antibody coding sequence (upon your request)

Applications

  • Provide accurate antibody sequencing to support and strengthen your patent application
  • Permanently store monoclonal antibody sequences and prevent the loss of hybridoma cell lines
  • Help the selection of appropriate cell lines to express recombinant monoclonal antibodies with higher purity
  • Shorten the period of hybridoma cloning and accelerate the generation of monoclonal antibodies
  • Affinity maturation further enhances the affinity of monoclonal antibody
  • Build chimeric antibodies, humanized antibodies, bi-specific antibodies, etc.