Hybridoma sequencing is generally based on reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technologies, which rapidly amplifies the 5’ and 3’ ends of cDNA from a low number of transcripts, and is widely used because it is simple, fast and inexpensive.First, we will extract your mRNA from your hybridoma cells and conduct RT-PCR to obtain the cDNA. Then, the light chain, heavy chain, or full-length fragment will be amplified by PCR or 3’ and 5’ RACE. The fragments can be connected to T-vectors and sequenced. First, we will extract your mRNA from your hybridoma cells and conduct RT-PCR to obtain the cDNA. Then, the light chain, heavy chain, or full-length fragment will be amplified by PCR or 3’ and 5’ RACE. The fragments can be connected to T-vectors and sequenced. 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.
Abvigen has years of experience in hybridoma sequencing. The sequencing range covers mice, rats, rabbits, goats, chickens, and other species. Abvigen has a mature antibody preparation platform, and also provides a recombinant antibody service.
Hybridoma cell line subclone Screening
Ensure that cells are monoclonal (prevent cell lines from being impure)
Detailed work performance report
Plasmid containing antibody gene sequence
Identification of hybridoma cell line subtypes
Determine the subtype to fit the appropriate primer
Amplification of antibody heavy chain and light chain
Amplification of antibody variable region (VH and VL only)
Complete sequence amplification of antibody (VH, VL, leading region and constant region)
Antibody sequence cloning
Cloning the amplified antibody sequence into T vector
Amplified antibody sequence gene sequencing
Antibody sequence analysis
Deduced amino acid sequence information of antibody according to antibody gene sequence
High-throughput + Great savings.
Fast turnaround time+ Reliability.
Comprehensive: Sequencing of heavy and light variable regions or full-length of antibody sequences.
Wide applicability: 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.
One-stop service: We provide a one-stop solution from hybridoma sequencing to recombinant antibody production.
After the two parties communicate and agree, determine the experimental plan, determine the service requirements - sign the contract - advance payment - start the experiment - deliver the results
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