A statistical approach to determine monoclonality after limiting cell plating of a hybridoma clone

J Immunol Methods. 1985 Feb 11;76(2):223-8. doi: 10.1016/0022-1759(85)90298-4.


One of the standard methods to isolate a hybridoma clone producing a monoclonal antibody requires successive steps of limiting dilution. The probability of obtaining a monoclonal antibody increases with the number of limiting dilution steps. However, without meticulous visual screening monoclonality is hard to prove. Here we describe a statistical analysis, based on Poisson's approximation, which allows one to calculate the number of hybridoma cells at a given plating efficiency so that when seeded a predictable number of mono-, bi-, etc.-clonal cultures are obtained after the first step of limiting cell plating.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / isolation & purification*
  • Hybridomas / cytology*
  • Mice
  • Mice, Inbred BALB C
  • Statistics as Topic


  • Antibodies, Monoclonal