Antibody Services

Our goal-driven custom antibody development program improves assay performance and reduces cost and timelines

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Monoclonal antibody development

We have the experience and technical expertise to meet the most challenging monoclonal antibody development goals. Our Ph.D. led hybridoma technical team works with customers from the design of a custom strategy and project plan, all the way through antibody development milestones. BBI scientists manage, analyse and communicate data, so you can make timely, responsive strategy decisions along the way.

Phase 1: Initiation

  • Hybridoma Initiation Checklist: To develop the best application specific antibody, communication is paramount. The Hybridoma Initiation Checklist is the customer’s first opportunity to provide us with the details necessary to strategise an optimal approach.
  • Technical Meeting: Open access to our technical resources and personnel begins with a conference call with the Hybridoma Development team to customise the project for success.

Phase 2: Immunisation

With a success rate of over 85%, the BBI Solutions Rapid Immunisation Protocol enables a fusion on day 28 of the project. This protocol ensures the fastest possible outcome for monoclonal antibody development.

  • Immunisation, Day 0: 3-5 mice are immunised with antigen. 2-3 mgs of antigen is required. 1 mg of additional counter-screening reagents required, if applicable.
  • Titre determination, Day 20: Titre is determined by indirect ELISA, additional assays can be performed. A mouse with adequate titre (1:31K) is selected for fusion. If titre is not reached by day 20, additional boosts may be administered.
  • Fusion boost, Day 24
  • Additional immunisation strategies include; subtractive immunisations (immunosupression), DNA immunisations, and cellular immunisations.

Phase 3: Fusion featuring MultiPure technology

  • Fusion, Day 28: The fused spleen(s) is plated across 10-20 plates (96 wells each). Fusion size is variable and is at the discretion of the customer.
  • Primary Screening, Day 38-42:Fusion product supernatants are screened by ELISA (primary screen) against the immunogen, along with any necessary counter-screens. ELISA options include: sandwich, cell based, bridging, blocking/inhibition, and indirect.
  • Scale-Up and Cryopreservation, approximately day 54: Depending on the amount of antibody required for customer evaluation, selected positive fusion products are scaled-up to generate 3-5 mLs of supernatant, or alternatively 15 mLs of overgrown supernatant. Supernatant from the 15 mL volume samples can be purified using our proprietary MultiPure technology. Two vials of each selected fusion product are cryopreserved at this stage.
  • Secondary Screening: Selected fusion products are screened a second time after scale up, by ELISA, to confirm stability and ensure active antibody secretion. A small percentage of fusion products can lose productivity during the scale-up phase and the secondary screen is important to confirm productivity before selecting fusion products for subcloning.
  • Once 15 mL supernatant samples are available, the customer can opt to use our novel MultiPure system to generate purified antibody. Doing so allows early insight into antibody performance, relative affinities, and even matched pair capabilities.  With the use of MultiPure, subcloning choices can be made with real, final application data saving both time and money.
  • Alternative fusion strategies can be discussed.
  • A fusion product(s) is selected for subcloning.

Phase 4: Subcloning

  • Subcloning: The selected fusion product(s) is subcloned by limiting dilution.
  • Subclone Screening: Screening by ELISA is performed against the immunogen and any necessary counter-screening agents.
  • Subclone, Scale-up and Cryopreservation: Approximately 2 weeks, depending on the amount of antibody required for  customer evaluation, selected subclones are scaled-up to generate 3-5 mLs of supernatant, or alternatively 15 mLs of overgrown supernatant. Supernatant from the 15 mL volume samples can be purified using the our MultiPure technology. Two vials of each selected subclone are cryopreserved at this stage.

Phase 5: Antibody production

Our premium monoclonal antibody development service is complimented by being a trusted partner in cGMP bulk antibody production.

  • In vitro production: In vitro production is performed in roller bottles. A 1 litre production takes approximately 3-4 weeks with an additional 1 week for purification. Production levels range from 20-50 mg/L depending on the cell line.
  • Yields are cell line dependent and are not guaranteed.

Discuss your requirements with our sales staff at (+1) 207-797-5454 or sales@bbisolutions.com

For more information about our polyclonal antibody development services, please visit our polyclonal antibody development section.

Learn more about our antibody production services

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