Hybridoma Screening Strategies

Having a high number of distinct positive hybridoma antibodies is a good outcome. However, a restricted budget often times means not all of them can be expanded, frozen, and the antibody purified and characterized. We believe an alternative to “brute force” production and characterization of positive hybridomas is the more precise selection based on secondary screening assays following an initial selection by solid-phase ELISA. The approach of using additional screening methods, or different antigens, often leads to several fit-for-purpose antibodies from a single fusion.

Advantages

  • Identify different antibody clones earlier in the clone selection part of the process
  • Know the apparent affinities of antibodies before production and purification
  • Select discrete antibodies to linear or conformational epitopes
  • Obtain metal-ion or cofactor dependent antibodies
  • Know relative antibody production levels of individual clones early in clone selection
 

Possible secondary screening methods include:

  • Blotting following SDS-PAGE and transfer to nitrocellulose, or directly using a dot blot apparatus
  • Quantitative analysis of kinetic rate constants and apparent affinity using Octet
  • Multiplex assays using fluorescent bead-based assays with different antigens bound to distinct bead sets
  • ELISA in the presence of metal ions or EDTA
  • Determination of isotype or IgG concentration to select high producing clones or a specific isotype antibody
  • COMING SOON! Clone selection using variable region sequencing