Hybridoma development services

Monoclonal Antibody form

Owing to a success rate of over 98% on more than 300 monoclonal antibody generations, ProteoGenix is able to offer the strongest guarantees on the market for hybridoma development. We are sharing this success with you by only requesting payment once your application is successful and has been handed over to you. Being able to offer this high level of guarantee makes us particularly proud!

Why choose ProteoGenix for
your hybridoma development ?

Guaranteed hybridoma development
Strongest guarantees

Test purified antibodies and only pay if you are satisfied! The best guarantees on the market!

Successful hybridoma generation
High success rate

More than 300 monoclonal antibody generation and over 98% success rate!

Animal immunization for antibody generation
Immunization approach diversity

Protein, peptide, DNA immunization… We carry out all types of immunization strategies!

aaalac hybridoma production
AAALAC accreditation

ProteoGenix applies the highest ethical standards and is committed to the ethical use of animals science.

Application guaranteed antibody
Screening in target application

Developing a diagnostic tool? Flow cytometry, Sandwich ELISA, IHC, IF, IP, WB… Your antibody is guaranteed in the application of your choice!

Therapeutic antibody expert
Therapeutic antibody expert

Looking for therapeutic antibody development? Reformatting (bispecific, ADC), humanization, affinity maturation… Use our wide range of services and go straight to the clinic!

Choose your guaranteed hybridoma development package

Which application do you need a monoclonal antibody for? Click on the corresponding application below to discover the content and guarantees of our hybridoma development packages.

Therapeutic antibody development
Therapeutic antibody development
IHF – Immunohistochemistry IHC guaranteed antibody services
Western Blot - WB guaranteed antibody services
Modification specific guaranteed hybridoma services
Immunoprecipitation - IP guaranteed antibody services
Immunofluorescence - IF guaranteed antibody services
Flow cytometry - FC guaranteed antibody services
ELISA guaranteed antibody services
Sandwich ELISA guaranteed hybridoma services
Package name Code Gene synthesis & antigen production Immunization, fusion, ELISA screening & subcloning Screening in application Purified Ab production Antibody conjugation & ELISA pair identification
Premium ELISA guaranteed ATX-PACK2M X X X
Premium WB guaranteed ATX-PACK3M X X X X
Premium Flow Cytometry guaranteed ATX-PACK4M X X X X
Premium ELISA Sandwich guaranteed ATX-PACK5M X X X X X
Premium IF guaranteed ATX-PACK6M X X X X
Premium IP guaranteed ATX-PACK7M X X X X
Peptide guaranteed ATX-PACK8M X X X
Modification specific guaranteed package ATX-PACK9M X X X
Premium IHC guaranteed ATX-PACK10M X X X

Hybridoma development testimonials

“We collaborated on a project for generating a monoclonal antibody that recognizes a one-amino-acid mutation in a cancer-associated cell-surface protein. Although the project was technically challenging, we succeeded in generating a hybridoma clone that produced the desired antibody. During the course of the project, we found additional clones that recognized adjacent epitopes and ProteoGenix proceeded to perform additional subclonings in order to ensure the delivery of clones that work perfectly. In summary, ProteoGenix always strived to ensure maximal product quality and customer satisfaction, and the account Manager provided prompt and competent support at all times.”

Satisfied monoclonal antibody customer

Dr. rer. nat. Rudolf Übelhart, Project Leader, Ulm Institute of Immunology, Germany

We congratulate the team of Dr. rer. nat. Ubelhart for their impressive achievements which lead to a publication in PNAS and we are proud that the antibodies generated by ProteoGenix contributed to their success!

“I requested ProteoGenix’s services for the generation of an anti-peptide monoclonal antibody to be used in my research. I required the antibody primarily for western blotting and immunoprecipitation experiments. Although the peptide antigen proved challenging in terms of immunogenicity, ProteoGenix was able to propose a range of customised strategies to overcome production issues encountered and finally succeeded in generating not only one but three antibodies of interest, thus offering me extra options in terms of final choice and use. During the whole project, Proteogenix strived to ensure maximal quality and to adapt to my needs, and my Account Manager always provided prompt and helpful services and solutions. I can highly commend both the level of expertise and the service I received.”

Dr. David Pryce, Lecturer in Biomedical Sciences (Immunology), Bangor University, UK

“We contacted ProteoGenix for a project concerning the detection of a phytopathogen. ProteoGenix helped us to the production of a specific antigen and the development of a monoclonal antibody to detect it by ELISA test and western blot. We appreciated the contact, the reactivity and professionalism of ProteoGenix for the good running of the project. We are satisfied with the services of the company.”

Université de Lille

Dr. Karine Lecointe, Researcher, Lille University, France

Our hybridoma development service content

Antigen design for hybridoma generation
  • Antigen design

Definition of the most relevant immunization strategy.
Antigen design: peptide synthesis, gene synthesis & protein production in 2 systems or DNA Immunization.

Immunization fro hybridoma development
  • Immunization

Immunization of 5 mice with our optimized proprietary protocol

cell fusion for hybridoma generation
  • Cell fusion

Collection of the splenocytes from 2 mice for 2 fusions with a myeloma cell line

Hybridoma selection
  • Hybridoma selection and screening (polyclonal stage)

Hybrid cells selection (HAT selection)
Culture supernatant screening vs. target antigen (ELISA screening)

Hybridoma cells expansion and selection
  • Isolation and selection of the best monoclones

Isolation of monoclones by limiting dilution.
Expansion and screening of the monoclones by ELISA or in target application.

Case study: monoclonal antibody production for the detection of a human cell membrane protein


A customer requested the production of a monoclonal antibody for the detection of a human cell membrane protein. Design of the antigen was particularly challenging as the protein was known to be difficult to express in mammalian cells.

Application guaranteed: Flow cytometry



  • A protein fragment estimated as the possible extra-cellular domain, and possible to express in mammalian cells, was designed.

  • The corresponding gene was designed and optimized for mammalian cell expression.

  • Mammalian cells were transfected with a plasmid expressing the target protein coupled to GFP.

Protein expression was performed both in HEK293 and in CHO cell lines. Final production was performed in HEK.

Production method: protein secreted in culture supernatant
Yield: 0.43mg/L
Quantity produced: 0.86mg
A final QC was performed by SDS-PAGE.


5 mice were immunized with 4 injections containing the protein antigen. More information about the protocol is available in the complete report.

FC results of the mice serum tested against the immunogen:

Sample tested Positive control Mouse 3 Mouse 4
Mice serum after 3rd injection 90% 42% 29%
Mice serum after 4th injection 76% 47%

Results for the other mice are available in the complete report.
Mouse 3 demonstrated the best immunogenic reactivity against the antigen in FC.
As our project includes 2 fusions, both mice 3 and 4 were selected to perform fusion.


Confirming screening after fusion
Clone ID 65 66 67 68 69 70 71
OD value 1.254 1.615 1.306 1.412 1.224 1.198 1.334
Clone ID 72 73 74 75 76 77 78
OD value 1.251 1.011 1.394 1.225 1.636 0.153 1.647
Clone ID 79 80 81 82 83 84 85
OD value 1.07 1.223 1.267 1.847 1.395 1.282 1.783
Clone ID 86 87 88 89 Negative Positive
OD value 1.036 0.983 1.173 1.116 0.088 1.731

24 positive clones were selected in ELISA.
Determination of the best clones for FC is done directly in the target application.



Details about the protocol are provided in the complete report.

Clone ID 65 66 67 68 69 70 71
FC result / % 37.48 1.27 20.27 62.50 1.18 11.40 1.38
Clone ID 72 73 74 75 76 77 78
FC result / % 75.09 1.01 6.50 56.76 73.16 1.18 1.09
Clone ID 79 80 81 82 83 84 85
FC result / % 1.07 1.05 1.49 6.75 71.92 1.18 1.02
Clone ID 86 87 88 Positive
FC result / % 1.04 1.01 7.36 80.73

11 positive clones were obtained after fusion in FC. These clones were subcloned for further antibody production.

Similar ELISA and FC analyses were performed on mouse 3. 14 positive clones were obtained after FC. Results are available in the complete report.



30 clones were selected by our customer for further production. Supernatant samples were tested in ELISA and FC after 2 subcloning steps. Results are provided in the complete report.



  • We succeeded in producing a difficult-to-express protein in sufficient quantities to develop monoclonal antibodies. Several expression tests, with several mammalian cells strains, were performed to reach this goal.

  • The immunization protocol in mice allowed us to develop good antibodies against the antigen.

  • After 2 fusions, 30 hybridomas/clones detecting the antigen in flow cytometry were produced.

  • 30 clones and 30 purified antibodies were delivered to our customer.


Please click on the button below to obtain the complete report.


Would you like to know more about our hybridoma generation services? Please take a look at another anti-peptide case study.

What are the advantages of hybridoma development for antibody generation?

Hybridoma technology revolutionized the whole biotechnological field by providing a method which is unlimited and reproducible for monoclonal antibody production. Today, hybridoma development is still a reference method for high sensitivity antibody generation. Therefore, hybridoma technology remains the best technique for assay development. It also represents a good alternative to phage display for therapeutic antibody development. However, administration of monoclonal antibodies produced by hybridoma technology leads to severe side effects due to their mouse origin. Thanks to recent advances in antibody engineering, the immunogenic mouse components can be reduced by a so-called antibody humanization process. Together with affinity maturation, this global process can be highly relevant to generate optimized humanized antibodies suitable for therapeutic applications.