Struggling with monoclonal antibody development? ProteoGenix’s custom hybridoma monoclonal antibody services, backed by over 20 years of expertise, offer a streamlined solution. Our fast-track protocol delivers high-affinity antibodies (up to 10^-12 M) in just 45 days, boasting a success rate exceeding 90% across 2,000+ projects and 20,000+ clones. We’re committed to your success, providing the strongest market guarantees, including payment only upon project completion. Our tailored services are designed to overcome challenges efficiently, making us the ideal partner for advancing your research. Contact ProteoGenix today for reliable, cost-effective solutions and transform your project challenges into success stories.

Different applications of our Custom Hybridoma Monoclonal Antibodies

Therapeutic Applications

ProteoGenix leverages over 20 years of expertise in hybridoma technology to develop potent antibody therapies with unmatched cost-efficiency. Our custom hybridoma development services produce antibodies with superior affinity, epitope specificity, and stability, ideal for challenging antigens. With 30+ antibodies in trials and 3 in the market, we’re your partner from lab to clinic. Discover more about our therapeutic antibody services.

Diagnostic as well as Research Applications

Start creating next-generation antibody-based diagnostics with ProteoGenix’s risk-free hybridoma production. Our custom hybridoma monoclonal antibody services guarantee optimal performance in any diagnostic application, including flow cytometry, Sandwich ELISA, IHC, IF, IP, and WB. With ten development packages and swift access to downstream services like antibody conjugation and custom ELISA development, we ensure your challenging projects swiftly move from concept to market. Our approach not only promises high-affinity antibodies tailored to your specific assay conditions and sample types but also empowers your diagnostic and research tools with unparalleled precision and reliability. Explore our comprehensive diagnostic antibody services to elevate your research and diagnostic capabilities.

Workflow of our Hybridoma development service

At ProteoGenix, we pride ourselves on over 20 years of expertise in delivering custom hybridoma monoclonal antibody services. Our process is designed to meet the highest standards of quality and efficiency, ensuring your research and therapeutic projects are supported by the best monoclonal antibodies available.

Antigen Design

  • Define immunization strategy
  • Perform peptide synthesis, gene synthesis, and protein production or opt for DNA immunization

Immunization

  • Immunize 5 mice using a proprietary protocol to generate a robust immune response

Cell Fusion

  • Collect splenocytes from 2 mice
  • Perform cell fusion with a myeloma cell line to create hybridomas

Hybridoma Selection And Screening

  • Select hybrid cells using HAT selection
  • Expand and screen monoclones by ELISA or in the target application for the highest quality antibodies

Isolation And Selection Of The Best Monoclones

  • Isolate monoclones through limiting dilution
  • Expand and screen monoclones by ELISA or in the target application for the highest quality antibodies

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

Aim Of The Project

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

Antigen Design And Production

  • 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.

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

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.

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.

Antibody Production

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.

Conclusion

  • 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.

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.”

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.”

“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.”

“I contacted ProteoGenix regarding the generation of monoclonal antibodies for the detection of a chemokine. ProteoGenix generated for us multiple monoclonal antibodies to detect the chemokine by ELISA, western blot and to test these antibodies in functional assays. They provided an excellent service, with regular updates, quick response to our queries and an excellent delivery of the product. I recommend their service for their efficacy and professionalism.”

“I highly recommend ProteoGenix for their fast and efficient services, providing top-quality antibodies suitable for diverse applications. Their exceptional communication, quick turnaround time, and reasonable pricing make them an excellent choice even for non-standard projects.”

“We contacted ProteoGenix to develop repertoires of specific antibodies against three antigens sharing high homology. As determined by ELISA, ProteoGenix developed several specific antibodies that exhibited no cross-reactivity with the other antigens. We appreciated their proactive approach to increase our chances of success (e.g., performing fusions at their own risks). We are satisfied with the services of the company.”

We needed to develop two monoclonal antibodies for Western-Blot, from antigen synthesis to hybridoma cell line and purified antibody delivery. ProteoGenix met all our expectations with efficient communication, satisfying deadlines, and successful production of specific and functional antibodies. For future antibody production, ProteoGenix will remain our preferred partner.“Dr. Simon Marlaire
Senior researcher at T3 Pharmaceuticals AG”

ProteoGenix’s Hybridoma platform (FAQ)

ProteoGenix Hybridoma Services FAQ

 

Hybridoma Antibody Screening

How are monoclonal Antibodies produced by Hybridoma

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