Monoclonal Antibodies_Mouse

Leinco Technologies offers over 25 years of demonstrated success in monoclonal antibody development for demanding applications such as early discovery research, in vivo preclinical therapeutic investigations, and in vitro diagnostics. Capabilities include antibodies developed in mice or Armenian hamsters against soluble proteins, cell surface molecules, or haptens for use in detection, neutralization or in vivo depletion. Utilizing proprietary development technologies, our scientific staff is skilled at producing monoclonal antibodies against single amino acid substitutions, splice variants and for neutralization.

Superior mAb Performance Using Mice or Hamsters

Leinco Technologies uniquely offers monoclonal antibody development using both mice or hamsters as host animals. Importantly, the use of hamster as the host animal offers the unique advantage of producing antibodies that can be used in vivo in mouse models without eliciting an immune response against the injected antibody.

Hybridoma Cell Line Development Process

Hybridoma cell line development consists of four (4) phases including immunization, fusion, cloning and in vitro antibody production. A protein or peptide-based project typically requires 4 – 6 mg of soluble antigen. Upon successful development of a panel of stable hybridoma cell lines, further characterization of secreted monoclonal antibodies and small-scale production is completed. In addition, at the conclusion of each project clients receive complete documentation of all phases and a development summary.

Phase Timeline Details
Phase 1: Immunization and Serum Screening 6-10 weeks
  • Immunize up to ten (10) mice or hamsters for each antigen using immunogens
  • Monitor antibody titer and/or neutralization activity, usually by ELISA
  • Determine the best animal(s) suited for B-cell fusions using screening data
Phase 2: Splenocyte Fusion and Screening of Parental Cultures 4-6 weeks
  • Splenocyte fusion of phase I animal selections
  • Splenocytes isolated and fused with myeloma cell lines
  • Fused cells under selection
  • Parental cultures from fusion screened
  • Stable positive cultures subcloned and expanded for cryopreservation
Phase 3: Subcloning and Screening 4-6 weeks

Cloning I:

  • 96 well panels screened
  • Positive clones selected for next round

Cloning II:

  • Second round of limiting dilution cloning
  • Stable clones expanded for further characterization
  • Supernatant from each clone sent to clients (optional)
Phase 4: In vitro Monoclonal Antibody Production 6-8 weeks
  • Complimentary in vitro production of approximately one liter of supernatant for one clone
  • Purification of supernatant using protein A chromatography and provided to client

Benefits of Partnering with Leinco

Communication with clients is of essential importance to quality monoclonal antibody development processes at Leinco Technologies . Our team of antibody experts can handle complex projects specific to clients’ needs while ensuring project goals are met. 

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