Entrust experienced professionals for antibody sequencing. Getting your monoclonal antibody sequence is a key starting point :

Our antibody sequencing process:
a one-stop solution from your hybridoma cell
line to your antibody sequence

Hybridoma cell line delivery

mRNA extraction

Reverse transcription

DNA sequencing of at least 5 samples for sequence alignment

Sub-cloning into a standard vector

PCR amplification

Data analysis and report writing

Optional: test expression + antibody/antigen interaction test

ProteoGenix offers a fully customizable antibody sequencing service with high guarantees. With 16 years of experience, our services are exhaustive and standardized and featured as follow for optimal performances:

  • Length of the sequence: antibody variable domains (VH, VL), full-length antibody (VH, VL, Fc)



  • A complete report including the CDR sequence annotated,
  • Plasmid containing the sequence (if requested),
  • Up to 2mg recombinant antibody (if requested).

Case study: DNA sequencing of variable
regions of hybridoma cell line

Aim Of The Study

Here is an example of a customer requesting to sequence a hybridoma cell line we had previously developed for him.


  • PacBio RS II
  • Hybridoma cells provided by ProteoGenix
  • TaKaRa MiniBEST Universal RNA Extraction Kit
  • PrimeScript RT reagent Kit with gDNA Eraser
  • TA-cloning Kit

Hybridoma Sequencing Protocol

Total RNA was extracted from hybridoma cells and cDNA subsequently synthesized. Antibody variable genes were then amplified by isotype-specific PCR, separately sub-cloned into a standard cloning vector and sequenced.

Isotype Determination And Antibody Sequence Analysis

Antibody isotype determination is performed by gel electrophoresis. Details of the protocol are described in the complete report provided in PDF file.

Results of the sequencing are based on the analysis of at least 10 positive clones and include:

  • Gene sequence
  • Amino acid sequence with annotated CDR

After completion of the antibody sequencing work, we were asked to engineer a chimeric version, synthesize both antibody cDNAs, subclone them into an expression vector, produce them in XtenCHO cells and determine antibody binding affinity (KD determination) of all 3 antibodies. The aim was to confirm that the chimeric recombinant antibody functionality at least remained equivalent to the mouse recombinant antibody and the mouse hybridoma antibody.

After confirmation, we were then able to develop humanized antibodies as well as bispecific antibodies for human therapeutic purposes.

Click on the button below to get the complete report “Parental Antibody Sequencing from Hybridoma”

Why should you sequence your antibody?

Patent application

  • The development of a monoclonal antibody can be the result of years of research. Demonstrating the uniqueness of your sequence is the first step to patenting it and securing your past and future investments.

Antibody engineering

  • Antibody sequencing provides valuable data for further engineering modifications.
  • For example, antibody humanization or affinity maturation can be performed once your mAb sequence has been obtained.
  • Also obtain access to various antibody formats such as bispecific antibodies or antibody-drug conjugates.

Recombinant production

  • Therapeutic antibody development requires a stable cell line able to produce a high amount of stable antibodies without contaminants.
  • Accessing your antibody sequence allows you to benefit from the advantages of recombinant antibody production: high reproducibility, ease of scalability and animal free production


  • Hybridoma cell lines can be lost due to gene loss, gene mutation or contamination.
  • Your hybridoma cell line sequence is the guarantee that you can continue recombinant antibody production even if your hybridoma line is lost.