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Purification and Characterisation

Antibody Purification 

Critical to antibody manufacture is the purification of specific and highly monomeric antibody. Our team of experts uses optimized protocols based on decades of experience to deliver purified antibody to meet your needs whether the source material is supernatant, ascites, or serum. 

We offer a wide range of *cGMP purification services. Antibody purification can be done as an extension of our monoclonal and polyclonal production offerings or as a stand-alone service. Based on years of purification experience with a vast number of cell culture supernatant, ascites and serum samples, our purification team has developed optimised protocols to generate high quality antibody in quantities sufficient for early screening studies through to large-scale production. 

Learn more about our available purification services: 

  • Protein A purification of mouse monoclonal cell lines with subclass-optimised binding and elution conditions 
  • Protein A and Protein G purification of polyclonal serum 
  • Antigen Affinity Purification of polyclonal antibodies using an immobilised antigen column
  • Low Endotoxin purification to generate purified antibody preparations with low EU/mg endotoxin concentrations 
  • MultiPure – library of purified hybridoma supernatants provided in a 96-well plate for early ranking assays 
  • ReadyPure – single milligram quantities of purified, subcloned mAb candidates for early characterisation and screening 

MultiPure Technology 

At the most critical juncture in a successful antibody development project, we enable our customers to efficiently and affordably select the most suitable clone for their application. By providing purified antibody earlier in the hybridoma development process, a MultiPure clone library allows for an unprecedented opportunity to generate data including specificity, relative affinities, matched pairs and more. 

Advantages 

  • Rapidly generates purified antibody from up to 94 fusion product supernatants at a time 
  • Early evaluation vastly improves the selection criteria of clones based on real data 
  • Normalisation of production rates allows for relative affinity comparison 
  • Additional plate of biotin conjugates allows for early matched-pair studies 

Technology 

Fusion products are expanded and supernatant is collected in up to 15ml quantities. The antibody is bound to a resin and eluted into a sealed 96 well plate. The typical yield for the process is 250ug-1mg per well. The range of concentrations seen with the MultiPure process is 0.3mg/ml-1.0mg/ml. Once purified, the antibody can be biotinylated and/or sterile filtered. Customers may also choose to add sodium azide to MultiPure samples. 

ReadyPure Technology 

In response to increasing customer interest in accessing small quantities of purified antibody samples earlier in the hybridoma development process, we developed the ReadyPure purification platform. Subcloned hybridoma supernatants are expanded to 100ml culture volumes, protein G purified, buffer exchanged and sterile filtered. ReadyPure offers customers 2-5mg average antibody yields to support more extensive screening and characterisation earlier in their project. Corresponding biotin conjugates can also be prepared for matched pair sandwich ELISA development. With over 27 years of hybridoma screening experience, we can design and execute early characterisation and assay development work in-house, saving you valuable time and resources. 

ReadyPure Highlights 

  • Hybridoma supernatant expanded to 100ml overgrown culture 
  • 2-5mg average antibody yields 
  • Purified antibody provided sterile filtered in PBS (standard) 
  • Custom final buffer formulations can be requested 
  • Biotin conjugates can be prepared 
  • ReadyPure samples can be shipped to the customer or maintained at our Maine facility for antibody characterisation and assay development work 

Characterisation 

Purified antibody can be characterised via: 

  • Monomeric purity via size exclusion chromatography (SEC) 
  • SDS-PAGE 
  • Isoelectric point via Isoelectric focussing (IEF) 
  • Zonal electrophoresis 
  • Western Blot 
  • Specificity via Enzyme-Linked Immunosorbent Assay (ELISA) 

Please contact our Customer Services Team directly to disuss your requirements on tel:+44 (0) 1495 363000 or alternatively you can submit an enquiry here.

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