Esterase amplification is the major organophosphorus (OP) insecticide resistance mechanism in Culex mosquitoes. The amplified Estalpha2(1)¿Estbeta2(1) esterases are found in > 90% of resistant populations worldwide, whereas amplified DNAs (amplicons) containing Estbeta1s are much rarer. Individuals with the Estbeta1 amplicons appear to be at a selective disadvantage in competition with those carrying the Estalpha2(1)¿Estbeta2(1) amplicons. To test the hypothesis that this is because Estbeta1 is less able to bind insecticide than the common amplified esterases, Estbeta1(2) was purified from the multi-resistant Habana strain of Culex quinquefasciatus, from Cuba. In its native form Estbeta1 is a monomeric enzyme of 66 kDa, with a pI of 4.8. The bimolecular rate constants for interaction of Estbeta1(2) with several OP insecticides were similar to those for the commonly elevated esterases Estalpha2(1) and Estbeta2(1), and much higher than for the electrophoretically identical non-elevated Estbeta1(3) and Estalpha3. Hence the apparent selective advantage of the Estalpha2(1)¿Estbeta2(1) amplicon is not due to its greater efficiency of insecticide binding, as OP insecticides are significantly better inhibitors of all the amplified esterases than of their non-amplified counterparts and therefore should be equally effective at conferring resistance.