Entrust experienced professionals for antibody sequencing. Getting your monoclonal antibody sequence is a key starting point :
- to protect you antibody sequence by patent application
- to optimize the potential of your antibody by further engineering
- to prevent the loss of the hybridoma cell line
- for recombinant antibody production.
Why choose ProteoGenix for your antibody sequencing?
Competitive price
Our prices are among the most competitive on the market!
No sequence no fee
If we do not succeed in determining the sequence? You don’t pay!
Fast turnaround
Benefit from a fast service thanks to our standardized process!
Monoclonal antibody development expert
Increase your success rate by using our engineering and production services!
Custom process
Get an antibody sequencing service strictly adapted to your requirements!
One stop solution
From early antibody discovery to the clinic!
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)
Optional:
- Gene synthesis with codon optimization for a chosen expression system: yeast, insect, mammalian
- Recombinant antibody production
- Interaction test against soluble antigen or cell antigen (ELISA, FACS,KD determination by Biacore / SPRi / Octet…)
Deliverables:
- 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.
Materials
- 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
- Safety
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.
What comes after your antibody
sequence analysis?
With more than 15 years of experience in antibody development and production, ProteoGenix can offer you far more than your hybridoma sequence. Using our antibody sequencing service can be the start of a long and fruitful collaboration that will significantly increase your chance to go to the clinic. ProteoGenix offers a one-stop solution tailored to take your project further, such as:
Therapeutic Antibody Development
ProteoGenix offers high-quality engineering solutions to optimize the properties of your therapeutic antibodies including antibody humanization, affinity maturation, bispecific antibodies, antibody drug conjugates etc..
Recombinant Antibody Production
ProteoGenix provides one of the highest performing recombinant antibody production platforms in the world thanks to 16 years of experience and proprietary technologies like XtenCHOTM.