Separation of anti-dinitrophenyl antibodies according to affinity by fractionated elution from immunoadsorbent using thiocyanate.

The possibility of fractionating anti-DNP-antibodies according to their affinity by successive elution from a DNP-immunoadsorbent with increasing concentrations of thiocyanate (KSCN) was investigated. Using KSCN at molarities of 0.5 M, 2.0 M and 3.0 M in a stepwise elution procedure, the mean affinity of the eluted antibody fractions was found to increase with increasing molarity of the eluent. The method described is a simple, reliable means of separating antibodies on an affinity basis which is of special value in systems where haptens are not available.